|
|
| Line 1: |
Line 1: |
| | {{Template:Cambridge2}}<!--Do not remove the first and last lines in this page!--> | | {{Template:Cambridge2}}<!--Do not remove the first and last lines in this page!--> |
| | | | |
| - | <html xmlns:v="urn:schemas-microsoft-com:vml"
| + | = Carotenoids = |
| | | | |
| - | xmlns:o="urn:schemas-microsoft-com:office:office"
| + | <!-- This is for the top grey / blue links bar !--> |
| | + | {{Template:Cambridgetemplatetop}} |
| | + | [https://2009.igem.org/Team:Cambridge/Project/Carotenoids/Int Introduction] |
| | + | [https://2009.igem.org/Team:Cambridge/Project/Carotenoids/PR Previous Research] |
| | + | [https://2009.igem.org/Team:Cambridge/Project/Carotenoids/OAP Our Action Plan] |
| | + | [https://2009.igem.org/Team:Cambridge/Project/Carotenoids/RA Result Analysis] |
| | + | [https://2009.igem.org/Team:Cambridge/Project/Carotenoids/Ref Reference] |
| | + | {{Template:Cambridgetemplatebottom}} |
| | | | |
| - | xmlns:w="urn:schemas-microsoft-com:office:word"
| + | == DESIGN OF DEVICES == |
| | + | === Biobricks contributed by previous iGEM teams === |
| | + | A few iGEM teams of previous years (e.g. Edinburgh 07, Edinburgh 08, Guelph 08) conducted research in the carotenoid system. We are fortunate to have access to some of their Biobricks in the registry: |
| | | | |
| - | xmlns:st1="urn:schemas-microsoft-com:office:smarttags"
| + | {| style="color:#CCC; background-color:#3D5089;" cellpadding="6" cellspacing="0" border="1" |
| | + | ! Registry Code |
| | + | ! Team |
| | + | ! Sequence Description |
| | + | ! Size of insert |
| | + | |- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1" |
| | + | |K118014 |
| | + | |Edinburgh 08 |
| | + | |[[Image:Icon translational unit.png]] (rbs + CrtE) |
| | + | |922bp |
| | + | |- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1" |
| | + | |K118006 |
| | + | |Edinburgh 08 |
| | + | |[[Image:Icon translational unit.png]] (rbs + CrtB) |
| | + | |949bp |
| | + | |- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1" |
| | + | |K118005 |
| | + | |Edinburgh 08 |
| | + | |[[Image:Icon translational unit.png]] (rbs + CrtI) |
| | + | |1498bp. 2 PstI sites removed. |
| | + | |- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1" |
| | + | |K118013 |
| | + | |Edinburgh 08 |
| | + | |[[Image:Icon translational unit.png]] (rbs + CrtY) |
| | + | |1162bp |
| | + | |- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1" |
| | + | |K152005 |
| | + | |Guelph 08 |
| | + | |[[Image:Icon translational unit.png]][[Image:Icon translational unit.png]][[Image:Icon translational unit.png]][[Image:Icon translational unit.png]][[Image:Icon reporter.png]] |
| | + | (rbs+CrtE) + (rbs+CrtB) + (rbs+CrtI) |
| | + | + (rbs+CrtY) + (rbs+GFP) |
| | + | |5412bp.2 PstI sites in CrtI removed. |
| | + | |- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1" |
| | + | |} |
| | | | |
| - | xmlns="http://www.w3.org/TR/REC-html40">
| |
| - |
| |
| - | <head>
| |
| - | <meta http-equiv=Content-Type content="text/html; charset=windows-1252">
| |
| - | <meta name=ProgId content=Word.Document>
| |
| - | <meta name=Generator content="Microsoft Word 11">
| |
| - | <meta name=Originator content="Microsoft Word 11">
| |
| - | <link rel=File-List href="Background.files/filelist.xml">
| |
| - | <link rel=Edit-Time-Data href="Background.files/editdata.mso">
| |
| - | <!--[if !mso]>
| |
| - | <style>
| |
| - | v\:* {behavior:url(#default#VML);}
| |
| - | o\:* {behavior:url(#default#VML);}
| |
| - | w\:* {behavior:url(#default#VML);}
| |
| - | .shape {behavior:url(#default#VML);}
| |
| - | </style>
| |
| - | <![endif]-->
| |
| - | <title>Background</title>
| |
| - | <o:SmartTagType namespaceuri="urn:schemas-microsoft-com:office:smarttags"
| |
| | | | |
| - | name="City"/>
| + | The above enzymes all come from a type of Gram negative Enterobacteria, Pantoea ananatis (GenBank: D90087.2). It is noted that analogues of these enzymes are also found in other organisms (e.g. bacteria Rhodobacter spp. and the fungus Neurospora crassa). |
| - | <o:SmartTagType namespaceuri="urn:schemas-microsoft-com:office:smarttags"
| + | |
| | | | |
| - | name="place"/>
| |
| - | <o:SmartTagType namespaceuri="urn:schemas-microsoft-com:office:smarttags"
| |
| | | | |
| - | name="chmetcnv"/>
| + | During our preliminary research, we encountered a few issues: |
| - | <!--[if gte mso 9]><xml>
| + | |
| - | <o:DocumentProperties>
| + | |
| - | <o:Author>Siming Ma</o:Author>
| + | |
| - | <o:Template>Normal</o:Template>
| + | |
| - | <o:LastAuthor>Siming Ma</o:LastAuthor>
| + | |
| - | <o:Revision>2</o:Revision>
| + | |
| - | <o:TotalTime>454</o:TotalTime>
| + | |
| - | <o:Created>2009-10-18T01:28:00Z</o:Created>
| + | |
| - | <o:LastSaved>2009-10-18T01:28:00Z</o:LastSaved>
| + | |
| - | <o:Pages>1</o:Pages>
| + | |
| - | <o:Words>2084</o:Words>
| + | |
| - | <o:Characters>11885</o:Characters>
| + | |
| - | <o:Lines>99</o:Lines>
| + | |
| - | <o:Paragraphs>27</o:Paragraphs>
| + | |
| - | <o:CharactersWithSpaces>13942</o:CharactersWithSpaces>
| + | |
| - | <o:Version>11.9999</o:Version>
| + | |
| - | </o:DocumentProperties>
| + | |
| - | </xml><![endif]--><!--[if gte mso 9]><xml>
| + | |
| - | <w:WordDocument>
| + | |
| - | <w:GrammarState>Clean</w:GrammarState>
| + | |
| - | <w:PunctuationKerning/>
| + | |
| - | <w:ValidateAgainstSchemas/>
| + | |
| - | <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid>
| + | |
| - | <w:IgnoreMixedContent>false</w:IgnoreMixedContent>
| + | |
| - | <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText>
| + | |
| - | <w:Compatibility>
| + | |
| - | <w:BreakWrappedTables/>
| + | |
| - | <w:SnapToGridInCell/>
| + | |
| - | <w:WrapTextWithPunct/>
| + | |
| - | <w:UseAsianBreakRules/>
| + | |
| - | <w:DontGrowAutofit/>
| + | |
| - | <w:UseFELayout/>
| + | |
| - | </w:Compatibility>
| + | |
| - | <w:BrowserLevel>MicrosoftInternetExplorer4</w:BrowserLevel>
| + | |
| - | </w:WordDocument>
| + | |
| - | </xml><![endif]--><!--[if gte mso 9]><xml>
| + | |
| - | <w:LatentStyles DefLockedState="false" LatentStyleCount="156">
| + | |
| - | </w:LatentStyles>
| + | |
| - | </xml><![endif]--><!--[if !mso]><object
| + | |
| - | classid="clsid:38481807-CA0E-42D2-BF39-B33AF135CC4D" id=ieooui></object>
| + | |
| - | <style>
| + | |
| - | st1\:*{behavior:url(#ieooui) }
| + | |
| - | </style>
| + | |
| - | <![endif]-->
| + | |
| - | <style>
| + | |
| - | <!--
| + | |
| - | /* Font Definitions */
| + | |
| - | @font-face
| + | |
| - | {font-family:Wingdings;
| + | |
| - | panose-1:5 0 0 0 0 0 0 0 0 0;
| + | |
| - | mso-font-charset:2;
| + | |
| - | mso-generic-font-family:auto;
| + | |
| - | mso-font-pitch:variable;
| + | |
| - | mso-font-signature:0 268435456 0 0 -2147483648 0;}
| + | |
| - | @font-face
| + | |
| - | {font-family:SimSun;
| + | |
| - | panose-1:2 1 6 0 3 1 1 1 1 1;
| + | |
| - | mso-font-alt:\5B8B\4F53;
| + | |
| - | mso-font-charset:134;
| + | |
| - | mso-generic-font-family:auto;
| + | |
| - | mso-font-pitch:variable;
| + | |
| - | mso-font-signature:3 135135232 16 0 262145 0;}
| + | |
| - | @font-face
| + | |
| - | {font-family:"Arial Black";
| + | |
| - | panose-1:2 11 10 4 2 1 2 2 2 4;
| + | |
| - | mso-font-charset:0;
| + | |
| - | mso-generic-font-family:swiss;
| + | |
| - | mso-font-pitch:variable;
| + | |
| - | mso-font-signature:647 0 0 0 159 0;}
| + | |
| - | @font-face
| + | |
| - | {font-family:\5B8B\4F53;
| + | |
| - | mso-font-alt:"Times New Roman";
| + | |
| - | mso-font-charset:0;
| + | |
| - | mso-generic-font-family:auto;
| + | |
| - | mso-font-pitch:variable;
| + | |
| - | mso-font-signature:0 0 0 0 0 0;}
| + | |
| - | @font-face
| + | |
| - | {font-family:"\@SimSun";
| + | |
| - | panose-1:2 1 6 0 3 1 1 1 1 1;
| + | |
| - | mso-font-charset:134;
| + | |
| - | mso-generic-font-family:auto;
| + | |
| - | mso-font-pitch:variable;
| + | |
| - | mso-font-signature:3 135135232 16 0 262145 0;}
| + | |
| - | /* Style Definitions */
| + | |
| - | p.MsoNormal, li.MsoNormal, div.MsoNormal
| + | |
| - | {mso-style-parent:"";
| + | |
| - | margin:0cm;
| + | |
| - | margin-bottom:.0001pt;
| + | |
| - | mso-pagination:widow-orphan;
| + | |
| - | font-size:12.0pt;
| + | |
| - | font-family:"Times New Roman";
| + | |
| - | mso-fareast-font-family:SimSun;}
| + | |
| - | h1
| + | |
| - | {mso-style-next:\6B63\6587;
| + | |
| - | margin-top:12.0pt;
| + | |
| - | margin-right:0cm;
| + | |
| - | margin-bottom:3.0pt;
| + | |
| - | margin-left:0cm;
| + | |
| - | mso-pagination:widow-orphan;
| + | |
| - | page-break-after:avoid;
| + | |
| - | mso-outline-level:1;
| + | |
| - | font-size:16.0pt;
| + | |
| - | font-family:Arial;
| + | |
| - | mso-font-kerning:16.0pt;
| + | |
| - | font-weight:bold;}
| + | |
| - | h2
| + | |
| - | {mso-style-next:\6B63\6587;
| + | |
| - | margin-top:12.0pt;
| + | |
| - | margin-right:0cm;
| + | |
| - | margin-bottom:3.0pt;
| + | |
| - | margin-left:0cm;
| + | |
| - | mso-pagination:widow-orphan;
| + | |
| - | page-break-after:avoid;
| + | |
| - | mso-outline-level:2;
| + | |
| - | font-size:14.0pt;
| + | |
| - | font-family:Arial;
| + | |
| - | font-weight:bold;
| + | |
| - | font-style:italic;}
| + | |
| - | h3
| + | |
| - | {mso-style-next:\6B63\6587;
| + | |
| - | margin-top:12.0pt;
| + | |
| - | margin-right:0cm;
| + | |
| - | margin-bottom:3.0pt;
| + | |
| - | margin-left:0cm;
| + | |
| - | mso-pagination:widow-orphan;
| + | |
| - | page-break-after:avoid;
| + | |
| - | mso-outline-level:3;
| + | |
| - | font-size:13.0pt;
| + | |
| - | font-family:Arial;
| + | |
| - | font-weight:bold;}
| + | |
| - | a:link, span.MsoHyperlink | + | |
| - | {color:blue;
| + | |
| - | text-decoration:underline;
| + | |
| - | text-underline:single;}
| + | |
| - | a:visited, span.MsoHyperlinkFollowed
| + | |
| - | {color:purple;
| + | |
| - | text-decoration:underline;
| + | |
| - | text-underline:single;}
| + | |
| - | p
| + | |
| - | {mso-margin-top-alt:auto;
| + | |
| - | margin-right:0cm;
| + | |
| - | mso-margin-bottom-alt:auto;
| + | |
| - | margin-left:0cm;
| + | |
| - | mso-pagination:widow-orphan;
| + | |
| - | font-size:12.0pt;
| + | |
| - | font-family:"Times New Roman";
| + | |
| - | mso-fareast-font-family:SimSun;}
| + | |
| - | span.mw-headline
| + | |
| - | {mso-style-name:mw-headline;}
| + | |
| - | span.apple-converted-space
| + | |
| - | {mso-style-name:apple-converted-space;}
| + | |
| - | span.apple-style-span
| + | |
| - | {mso-style-name:apple-style-span;}
| + | |
| - | p.TabellenInhalt, li.TabellenInhalt, div.TabellenInhalt
| + | |
| - | {mso-style-name:"Tabellen Inhalt";
| + | |
| - | margin:0cm;
| + | |
| - | margin-bottom:.0001pt;
| + | |
| - | mso-pagination:no-line-numbers;
| + | |
| - | mso-hyphenate:none;
| + | |
| - | font-size:12.0pt;
| + | |
| - | font-family:"Times New Roman";
| + | |
| - | mso-fareast-font-family:\5B8B\4F53;
| + | |
| - | mso-font-kerning:.5pt;
| + | |
| - | mso-fareast-language:#00FF;}
| + | |
| - | span.GramE
| + | |
| - | {mso-style-name:"";
| + | |
| - | mso-gram-e:yes;}
| + | |
| - | @page Section1
| + | |
| - | {size:612.0pt 792.0pt;
| + | |
| - | margin:9.0pt 90.0pt 18.0pt 90.0pt;
| + | |
| - | mso-header-margin:35.4pt;
| + | |
| - | mso-footer-margin:35.4pt;
| + | |
| - | mso-paper-source:0;}
| + | |
| - | div.Section1
| + | |
| - | {page:Section1;}
| + | |
| - | /* List Definitions */
| + | |
| - | @list l0
| + | |
| - | {mso-list-id:120536249;
| + | |
| - | mso-list-type:hybrid;
| + | |
| - | mso-list-template-ids:2126427888 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| + | |
| - | @list l0:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0B7;
| + | |
| - | mso-level-tab-stop:36.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | font-family:Symbol;}
| + | |
| - | @list l1
| + | |
| - | {mso-list-id:329873205;
| + | |
| - | mso-list-type:hybrid;
| + | |
| - | mso-list-template-ids:422478762 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| + | |
| - | @list l1:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0B7;
| + | |
| - | mso-level-tab-stop:36.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | font-family:Symbol;}
| + | |
| - | @list l2
| + | |
| - | {mso-list-id:449712540;
| + | |
| - | mso-list-template-ids:249083244;}
| + | |
| - | @list l2:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0A7;
| + | |
| - | mso-level-tab-stop:36.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | mso-ansi-font-size:10.0pt;
| + | |
| - | font-family:Wingdings;}
| + | |
| - | @list l3
| + | |
| - | {mso-list-id:632642131;
| + | |
| - | mso-list-template-ids:-220050230;}
| + | |
| - | @list l3:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0A7;
| + | |
| - | mso-level-tab-stop:36.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | mso-ansi-font-size:10.0pt;
| + | |
| - | font-family:Wingdings;}
| + | |
| - | @list l4
| + | |
| - | {mso-list-id:1088690536;
| + | |
| - | mso-list-type:hybrid;
| + | |
| - | mso-list-template-ids:-465797406 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| + | |
| - | @list l4:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0B7;
| + | |
| - | mso-level-tab-stop:72.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | margin-left:72.0pt;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | font-family:Symbol;}
| + | |
| - | @list l5
| + | |
| - | {mso-list-id:1133330608;
| + | |
| - | mso-list-type:hybrid;
| + | |
| - | mso-list-template-ids:1158428478 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| + | |
| - | @list l5:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0B7;
| + | |
| - | mso-level-tab-stop:36.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | font-family:Symbol;}
| + | |
| - | @list l6
| + | |
| - | {mso-list-id:1410152836;
| + | |
| - | mso-list-template-ids:-1665133950;}
| + | |
| - | @list l6:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0A7;
| + | |
| - | mso-level-tab-stop:36.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | mso-ansi-font-size:10.0pt;
| + | |
| - | font-family:Wingdings;}
| + | |
| - | @list l7
| + | |
| - | {mso-list-id:2013410533;
| + | |
| - | mso-list-type:hybrid;
| + | |
| - | mso-list-template-ids:-248105022 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| + | |
| - | @list l7:level1
| + | |
| - | {mso-level-number-format:bullet;
| + | |
| - | mso-level-text:\F0B7;
| + | |
| - | mso-level-tab-stop:18.0pt;
| + | |
| - | mso-level-number-position:left;
| + | |
| - | margin-left:18.0pt;
| + | |
| - | text-indent:-18.0pt;
| + | |
| - | font-family:Symbol;}
| + | |
| - | ol
| + | |
| - | {margin-bottom:0cm;}
| + | |
| - | ul
| + | |
| - | {margin-bottom:0cm;}
| + | |
| - | -->
| + | |
| - | </style>
| + | |
| - | <!--[if gte mso 10]>
| + | |
| - | <style>
| + | |
| - | /* Style Definitions */
| + | |
| - | table.MsoNormalTable
| + | |
| - | {mso-style-name:\666E\901A\8868\683C;
| + | |
| - | mso-tstyle-rowband-size:0;
| + | |
| - | mso-tstyle-colband-size:0;
| + | |
| - | mso-style-noshow:yes;
| + | |
| - | mso-style-parent:"";
| + | |
| - | mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
| + | |
| - | mso-para-margin:0cm;
| + | |
| - | mso-para-margin-bottom:.0001pt;
| + | |
| - | mso-pagination:widow-orphan;
| + | |
| - | font-size:10.0pt;
| + | |
| - | font-family:"Times New Roman";
| + | |
| - | mso-ansi-language:#0400;
| + | |
| - | mso-fareast-language:#0400;
| + | |
| - | mso-bidi-language:#0400;}
| + | |
| - | table.MsoTableGrid
| + | |
| - | {mso-style-name:\7F51\683C\578B;
| + | |
| - | mso-tstyle-rowband-size:0;
| + | |
| - | mso-tstyle-colband-size:0;
| + | |
| - | border:solid windowtext 1.0pt;
| + | |
| - | mso-border-alt:solid windowtext .5pt;
| + | |
| - | mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
| + | |
| - | mso-border-insideh:.5pt solid windowtext;
| + | |
| - | mso-border-insidev:.5pt solid windowtext;
| + | |
| - | mso-para-margin:0cm;
| + | |
| - | mso-para-margin-bottom:.0001pt;
| + | |
| - | mso-pagination:widow-orphan;
| + | |
| - | font-size:10.0pt;
| + | |
| - | font-family:"Times New Roman";
| + | |
| - | mso-ansi-language:#0400;
| + | |
| - | mso-fareast-language:#0400;
| + | |
| - | mso-bidi-language:#0400;}
| + | |
| - | </style>
| + | |
| - | <![endif]--><!--[if gte mso 9]><xml>
| + | |
| - | <o:shapedefaults v:ext="edit" spidmax="2050"/>
| + | |
| - | </xml><![endif]--><!--[if gte mso 9]><xml>
| + | |
| - | <o:shapelayout v:ext="edit">
| + | |
| - | <o:idmap v:ext="edit" data="1"/>
| + | |
| - | </o:shapelayout></xml><![endif]-->
| + | |
| - | </head>
| + | |
| - |
| + | |
| - | <body lang=EN-US link=blue vlink=purple style='tab-interval:36.0pt'>
| + | |
| - |
| + | |
| - | <div class=Section1>
| + | |
| - |
| + | |
| - | <h1 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| + | |
| | | | |
| - | 0cm'>BACKGROUND</h1>
| + | * Originally, the registry annotation for K152005 indicated that there was no ribosome binding site before gene CrtI (i.e. “…rbs+CrtB+CrtI+rbs+CrtY…”). After clarification with Team Guelph 08, we were told that it was an error in annotation. The registry entry for K152005 has since been updated to include the ribosome binding site. |
| - |
| + | |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| + | |
| | | | |
| - | 0cm;text-align:justify;text-justify:inter-ideograph;line-height:150%'><span
| + | * Teams of previous years constructed various composite parts containing two or more enzymes of the synthetic pathways (e.g. CrtEBI, CrtEBIY), but they are not available on the 2009 Distribution Plates. |
| | | | |
| - | style='font-size:11.0pt;line-height:150%;font-weight:normal;mso-bidi-font-weight:
| + | * We put K152005 under control of constitutive promoter (R0011) and transformed the construct into E.coli TOP10 cells. However, the plasmids seemed unstable (the colonies streaks on solid agar plates were inconsistent in colour) and the pigment production was low. This indicated that a different strain of E.coli might be needed for pigment production. |
| | | | |
| - | bold'>Carotenoids are organic pigments naturally present in plants, algae and
| |
| - | some bacteria. There are more than 600 carotenoids, which can be categorized
| |
| - | into xanthophylls (hydrocarbons containing oxygen) and carotenes (hydrocarbons
| |
| - | containing no oxygen). Carotenoids perform a range of functions, including
| |
| - | light energy absorption, protection against photo-damage, acting as
| |
| - | antioxidants, and as precursor to other organic compounds. In human, for
| |
| - | example, beta-carotene is the precursor to vitamin A.<o:p></o:p></span></h3>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| | | | |
| - | 0cm'><u>Biochemical pathway of carotenoid synthesis<o:p></o:p></u></h3>
| + | === Create our own devices by standard assembly === |
| - |
| + | Given the information at hand, we decided to adopt the following general strategies: |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| + | |
| | | | |
| - | 0cm;text-align:justify;text-justify:inter-ideograph;line-height:150%'><span
| + | * Create a lycopene-producing device (with enzymes CrtE, CrtB and CrtI) by standard assembly of Biobricks K118014, K118006 and K118005. |
| | | | |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial'>The common starting
| + | * Create a β-carotene-producing device (with enzymes CrtE, CrtB, CrtI and CrtY) by standard assembly of Biobricks K118014, K118006, K118005 and K118013. |
| - | point for carotenoid synthesis is<span class=apple-converted-space> </span><span
| + | |
| | | | |
| - | style='mso-bidi-font-weight:bold'>farnesyl pyrophosphate (FPP)</span>, which
| + | * Put the above two constructs under constitutive promoter (R0011) and arabinose-induced promoter/Pbad (I0500) and test the effect on colour production. |
| - | derives from two precursors, isopentenyl pyrophosphate (IPP) and dimethylallyl
| + | |
| - | pyrophosphate (DMAPP). In general, there are two pathways for synthesising IPP
| + | |
| - | and DMAPP: the Mevalonate Pathway (starting with acetyl CoA) and the | + | |
| - | Non-mevalonate Pathway (starting with pyruvate and glyceradehyde-3-phosphate).
| + | |
| - | While the Mevalonate Pathway is present in all higher eukaryotes, the<span
| + | |
| | | | |
| - | class=apple-converted-space> </span><span style='mso-bidi-font-weight:
| |
| | | | |
| - | bold'>Non-mevalonate Pathway</span><span class=apple-converted-space> </span>is
| |
| - | present in<span class=apple-converted-space> </span><i>E. coli</i>.<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
| |
| - | <td width=307 valign=top style='width:230.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center;background:#F9F9F9'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Cam09_Mevalonate_pathway.png"
| |
| - |
| |
| - | title="Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)"><!--[if gte vml 1]><v:shapetype
| |
| - | id="_x0000_t75" coordsize="21600,21600" o:spt="75" o:preferrelative="t"
| |
| - | path="m@4@5l@4@11@9@11@9@5xe" filled="f" stroked="f">
| |
| - | <v:stroke joinstyle="miter"/>
| |
| - | <v:formulas>
| |
| - | <v:f eqn="if lineDrawn pixelLineWidth 0"/>
| |
| - | <v:f eqn="sum @0 1 0"/>
| |
| - | <v:f eqn="sum 0 0 @1"/>
| |
| - | <v:f eqn="prod @2 1 2"/>
| |
| - | <v:f eqn="prod @3 21600 pixelWidth"/>
| |
| - | <v:f eqn="prod @3 21600 pixelHeight"/>
| |
| - | <v:f eqn="sum @0 0 1"/>
| |
| - | <v:f eqn="prod @6 1 2"/>
| |
| - | <v:f eqn="prod @7 21600 pixelWidth"/>
| |
| - | <v:f eqn="sum @8 21600 0"/>
| |
| - | <v:f eqn="prod @7 21600 pixelHeight"/>
| |
| - | <v:f eqn="sum @10 21600 0"/>
| |
| - | </v:formulas>
| |
| - | <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>
| |
| - | <o:lock v:ext="edit" aspectratio="t"/>
| |
| - | </v:shapetype><v:shape id="_x0000_i1025" type="#_x0000_t75" alt="Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)"
| |
| - | title=""Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)""
| |
| - | style='width:214.5pt;height:132.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image001.png" o:href="https://static.igem.org/mediawiki/2009/thumb/d/d1/Cam09_Mevalonate_pathway.png/334px-Cam09_Mevalonate_pathway.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=286 height=177
| |
| - |
| |
| - | src="Background.files/image002.gif"
| |
| - |
| |
| - | alt="Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)"
| |
| - |
| |
| - | v:shapes="_x0000_i1025"><![endif]></a><o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><b><span style='font-size:8.5pt;font-family:Arial'>Mevalonate Pathway</span></b><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:8.5pt;font-family:Arial'> </span></span><span
| |
| - |
| |
| - | style='font-size:8.5pt;font-family:Arial'>(image adopted from Wikipedia, <a
| |
| - |
| |
| - | href="http://en.wikipedia.org/wiki/Mevalonate_pathway"
| |
| - |
| |
| - | title="http://en.wikipedia.org/wiki/Mevalonate_pathway"><span
| |
| - |
| |
| - | style='color:windowtext;text-decoration:none;text-underline:none'>http://en.wikipedia.org/wiki/Mevalonate_pathway</span></a>)</span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=283 valign=top style='width:212.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:9.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Cam09_Non_mevalonate.JPG"
| |
| - |
| |
| - | title="Non-mevalonate Pathway (image adopted from Yuan et al. 2006)"><span
| |
| - |
| |
| - | style='color:windowtext;text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1026" type="#_x0000_t75" alt="Non-mevalonate Pathway (image adopted from Yuan et al. 2006)"
| |
| - | title=""Non-mevalonate Pathway (image adopted from Yuan et al. 2006)""
| |
| - | style='width:201.75pt;height:141pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image003.jpg" o:href="https://static.igem.org/mediawiki/2009/thumb/1/19/Cam09_Non_mevalonate.JPG/360px-Cam09_Non_mevalonate.JPG"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=269 height=188
| |
| - |
| |
| - | src="Background.files/image004.jpg"
| |
| - |
| |
| - | alt="Non-mevalonate Pathway (image adopted from Yuan et al. 2006)" v:shapes="_x0000_i1026"><![endif]></span></a></span><b><span
| |
| - |
| |
| - | style='font-size:8.5pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>Non-mevalonate Pathway</span></b><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:8.5pt;font-family:Arial'> </span></span><span
| |
| - |
| |
| - | style='font-size:8.5pt;font-family:Arial'>(image adopted from Yuan et al.
| |
| - | 2006)</span><span style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;text-align:justify;text-justify:inter-ideograph'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'><br>
| |
| - | </span><span style='font-size:11.0pt;font-family:Arial'>After IPP and DMAPP go
| |
| - | on to form FPP, a series of enzymatic reactions convert the colourless FPP to
| |
| - | coloured compounds: red lycopene, and orange β-carotene.<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
| |
| - | <td width=590 valign=top style='width:442.8pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1027" type="#_x0000_t75" style='width:6in;height:282.75pt'>
| |
| - | <v:imagedata src="Background.files/image005.jpg" o:title="Cambridge_Carotenoid_100"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=576 height=377
| |
| - |
| |
| - | src="Background.files/image006.jpg" v:shapes="_x0000_i1027"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span class=apple-style-span><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial;mso-bidi-font-weight:bold'>By modulating the enzymes
| |
| - | involved in this conversion pathway ("CrtE", "CrtB",
| |
| - | "CrtI", and "CrtY"), we aim to produce and control
| |
| - | different pigments as signal output.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <b><span style='font-size:16.0pt;font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | SimSun;mso-font-kerning:16.0pt;mso-ansi-language:EN-US;mso-fareast-language:
| |
| - |
| |
| - | ZH-CN;mso-bidi-language:AR-SA'><br clear=all style='page-break-before:always'>
| |
| - | </span></b>
| |
| - |
| |
| - | <h1 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'>DESIGN OF DEVICES</h1>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><u>Biobricks contributed by previous iGEM teams<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>A few iGEM
| |
| - | teams of previous years (e.g. <st1:City w:st="on">Edinburgh</st1:City> 07, <st1:City
| |
| - |
| |
| - | w:st="on">Edinburgh</st1:City> 08, <st1:place w:st="on"><st1:City w:st="on"><span
| |
| - |
| |
| - | class=GramE>Guelph</span></st1:City></st1:place> 08) conducted research in
| |
| - | the carotenoid system. We are fortunate to have access to some of their
| |
| - | Biobricks in the registry:<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='mso-cellspacing:0cm;background:#3D5089;mso-padding-alt:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=67 style='width:50.55pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Registry Code<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Team<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Sequence Description<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Size of insert<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118014<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1028"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1028"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtE)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>922bp<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118006<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1029"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1029"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtB)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>949bp<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:3'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118005<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1030"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1030"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtI)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>1498bp<o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'>2 PstI sites removed.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:4'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118013<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1031"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1031"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtY)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>1162bp<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K152005<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Guelph</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1032"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1032"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1033"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1033"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1034"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1034"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1035"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1035"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_reporter.png" title="Image:Icon reporter.png"><span
| |
| - |
| |
| - | style='color:windowtext;text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1036" type="#_x0000_t75" alt="Image:Icon reporter.png" title=""Image:Icon reporter.png""
| |
| - | style='width:27.75pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image009.png" o:href="https://static.igem.org/mediawiki/2009/a/aa/Icon_reporter.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=37 height=25
| |
| - |
| |
| - | src="Background.files/image010.jpg" alt="Image:Icon reporter.png" v:shapes="_x0000_i1036"><![endif]></span></a><o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'>(rbs+CrtE) + (rbs+CrtB)
| |
| - | + (rbs+CrtI) + (rbs+CrtY) + (rbs+GFP)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>5412bp<o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'>2 PstI sites in CrtI
| |
| - | removed.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span class=apple-converted-space><span style='font-size:
| |
| - |
| |
| - | 11.0pt;mso-bidi-font-size:12.0pt;line-height:150%;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>The above enzymes all come from a type of Gram negative
| |
| - | Enterobacteria, <i style='mso-bidi-font-style:normal'>Pantoea ananatis</i> (GenBank:
| |
| - | D90087.2). It is noted that analogues of these enzymes are also found in other
| |
| - | organisms (e.g. bacteria <i style='mso-bidi-font-style:normal'>Rhodobacter</i>
| |
| - | spp. and the fungus <i style='mso-bidi-font-style:normal'>Neurospora crassa</i>).<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span class=apple-converted-space><span style='font-size:
| |
| - |
| |
| - | 11.0pt;mso-bidi-font-size:12.0pt;line-height:150%;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>During our preliminary research, we have encountered a few
| |
| - | issues:<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:35.7pt;text-indent:-17.85pt;mso-list:l5 level1 lfo6;tab-stops:list 36.0pt'><![if !supportLists]><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:11.0pt;mso-bidi-font-size:
| |
| - |
| |
| - | 12.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
| |
| - |
| |
| - | Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>
| |
| - | </span></span></span></span><![endif]><span class=apple-converted-space><span
| |
| - |
| |
| - | style='font-size:11.0pt;mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>Originally, the registry annotation for K152005 indicated
| |
| - | that there was no ribosome binding site before gene CrtI (i.e.
| |
| - | “…rbs+CrtB+CrtI+rbs+CrtY…”). After clarification with Team Guelph 08, we were
| |
| - | told that it was an error in annotation. The registry entry for K152005 has
| |
| - | since been updated to include the ribosome binding site.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:35.7pt;text-indent:-17.85pt;mso-list:l5 level1 lfo6;tab-stops:list 36.0pt'><![if !supportLists]><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:11.0pt;mso-bidi-font-size:
| |
| - |
| |
| - | 12.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
| |
| - |
| |
| - | Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>
| |
| - | </span></span></span></span><![endif]><span class=apple-converted-space><span
| |
| - |
| |
| - | style='font-size:11.0pt;mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>Teams of previous years constructed various composite parts
| |
| - | containing two or more enzymes of the synthetic pathways (e.g. CrtEBI,
| |
| - | CrtEBIY), but they are not available on the 2009 Distribution Plates.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:35.7pt;text-indent:-17.85pt;mso-list:l5 level1 lfo6;tab-stops:list 36.0pt'><![if !supportLists]><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:11.0pt;mso-bidi-font-size:
| |
| - |
| |
| - | 12.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
| |
| - |
| |
| - | Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>
| |
| - | </span></span></span></span><![endif]><span class=apple-converted-space><span
| |
| - |
| |
| - | style='font-size:11.0pt;mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>We put K152005 under control of constitutive promoter
| |
| - | (R0011) and transformed the construct into E.coli TOP10 cells. However, the
| |
| - | plasmids seemed unstable (the colonies streaks on solid agar plates were
| |
| - | inconsistent in colour) and the pigment production was low. This indicated that
| |
| - | a different strain of E.coli might be needed for pigment production.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><span class=apple-converted-space><span style='font-size:11.0pt;
| |
| - |
| |
| - | mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></span></p>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><u>Create our own devices by standard assembly<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><span style='font-size:11.0pt;font-family:Arial'>Given the
| |
| - | information at hand, we decided to adopt the following general strategies:<o:p></o:p></span></p>
| |
| - |
| |
| - | <ul style='margin-top:0cm' type=disc>
| |
| - | <li class=MsoNormal style='margin-top:6.0pt;margin-bottom:6.0pt;mso-list:l0 level1 lfo7;
| |
| - |
| |
| - | tab-stops:list 36.0pt'><span style='font-size:11.0pt;font-family:Arial'>Create
| |
| - | a lycopene-producing device (with enzymes CrtE, CrtB and CrtI) by standard
| |
| - | assembly of Biobricks K118014, K118006 and K118005.<o:p></o:p></span></li>
| |
| - | <li class=MsoNormal style='margin-top:6.0pt;margin-bottom:6.0pt;mso-list:l0 level1 lfo7;
| |
| - |
| |
| - | tab-stops:list 36.0pt'><span style='font-size:11.0pt;font-family:Arial'>Create
| |
| - | a <span style='mso-bidi-font-weight:bold'>β-carotene-producing device
| |
| - | (with enzymes CrtE, CrtB, CrtI and CrtY) by standard assembly of Biobricks
| |
| - | K118014, K118006, K118005 and K118013.</span><o:p></o:p></span></li>
| |
| - | <li class=MsoNormal style='margin-top:6.0pt;margin-bottom:6.0pt;mso-list:l0 level1 lfo7;
| |
| - |
| |
| - | tab-stops:list 36.0pt'><span style='font-size:11.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-weight:bold'>Put the above two constructs under constitutive
| |
| - | promoter (R0011) and arabinose-induced promoter/Pbad (I0500) and test the
| |
| - | effect on colour production.</span><span style='font-size:11.0pt;
| |
| - |
| |
| - | font-family:Arial'><o:p></o:p></span></li>
| |
| - | </ul>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:11.0pt'>Scheme for standard assembly<o:p></o:p></span></h2>
| |
| - |
| |
| - | <div align=center>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=90 valign=top style='width:67.5pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial;mso-fareast-font-family:Arial'>Lycopene-producing
| |
| - | device<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b><span style='font-size:10.0pt;font-family:Arial'>β-carotene-producing
| |
| - | device</span></b><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial;mso-fareast-font-family:Arial'><o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Constituent
| |
| - | Biobricks (already in the registry)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1037"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropright="41936f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image012.jpg" v:shapes="_x0000_i1037"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118014)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1038"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="21948f" cropright="21948f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image013.jpg" v:shapes="_x0000_i1038"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118006)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1039"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="42249f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image014.jpg" v:shapes="_x0000_i1039"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118005)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1040"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropright="41936f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image012.jpg" v:shapes="_x0000_i1040"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118014)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1041"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="21948f" cropright="21948f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image013.jpg" v:shapes="_x0000_i1041"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118006)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1042"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="42249f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image014.jpg" v:shapes="_x0000_i1042"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118005)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1043"
| |
| - | type="#_x0000_t75" style='width:59.25pt;height:24pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"
| |
| - | cropbottom="-2030f" cropleft="50400f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=79 height=32
| |
| - |
| |
| - | src="Background.files/image016.jpg" v:shapes="_x0000_i1043"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118013)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Basic construct<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1044" type="#_x0000_t75"
| |
| - | style='width:135pt;height:24pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=180 height=32
| |
| - |
| |
| - | src="Background.files/image017.jpg" v:shapes="_x0000_i1044"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial'>K274100</span></b><span style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1045" type="#_x0000_t75"
| |
| - | style='width:160.5pt;height:24pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"
| |
| - | cropbottom="-2114f" cropleft="9882f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=214 height=32
| |
| - |
| |
| - | src="Background.files/image018.jpg" v:shapes="_x0000_i1045"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial'>K274200</span></b><span style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:3'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Under constitutive promoter<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1046" type="#_x0000_t75"
| |
| - | style='width:155.25pt;height:26.25pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"
| |
| - | cropbottom="-2427f" cropright="13653f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=207 height=35
| |
| - |
| |
| - | src="Background.files/image019.jpg" v:shapes="_x0000_i1046"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial;mso-fareast-font-family:Arial'>K274110</span></b><span
| |
| - |
| |
| - | style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1047" type="#_x0000_t75"
| |
| - | style='width:170.25pt;height:23.25pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=227 height=31
| |
| - |
| |
| - | src="Background.files/image020.jpg" v:shapes="_x0000_i1047"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial;mso-fareast-font-family:Arial'>K274210</span></b><span
| |
| - |
| |
| - | style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Under Pbad promoter<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1048" type="#_x0000_t75"
| |
| - | style='width:157.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image021.png" o:title="untitled3"
| |
| - | cropright="12765f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=210 height=33
| |
| - |
| |
| - | src="Background.files/image022.jpg" v:shapes="_x0000_i1048"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial;mso-fareast-font-family:Arial'>K274120</span></b><span
| |
| - |
| |
| - | style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1049" type="#_x0000_t75"
| |
| - | style='width:170.25pt;height:23.25pt'>
| |
| - | <v:imagedata src="Background.files/image021.png" o:title="untitled3"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=227 height=31
| |
| - |
| |
| - | src="Background.files/image023.jpg" v:shapes="_x0000_i1049"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial'>K274220</span></b><span style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span style='font-size:11.0pt;line-height:150%;
| |
| - |
| |
| - | font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span style='font-size:11.0pt;line-height:150%;
| |
| - |
| |
| - | font-family:Arial;mso-bidi-font-family:"Times New Roman"'>After constructing
| |
| - | our devices, we conducted restriction digest (cut with enzymes XbaI and PstI on
| |
| - | Biobrick prefix and suffix, respectively) to check that the sizes of the
| |
| - | inserts were correct. The results are as follows:<o:p></o:p></span></p>
| |
| - |
| |
| - | <div align=center>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:24.55pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Biobrick Number<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Size of insert (kb)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Size of backbone (kb)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Note<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;height:7.6pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274100<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>3.37<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Digestion
| |
| - | result not shown.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2;height:8.05pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274110<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>3.42<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:3;height:17.55pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274120<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.58<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.43<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Sizes
| |
| - | of insert and backbone were similar so only one band appeared on gel.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:4;height:8.95pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274200<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.53<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Digestion
| |
| - | result not shown.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:5;height:8.1pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274210<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.59<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:6;height:8.15pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274220<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>5.74<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.43<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:7;height:23.25pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'><span style='font-size:10.0pt;font-family:
| |
| - |
| |
| - | Arial'>R0011-K152005*<o:p></o:p></span></i></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>5.48<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>2.08<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | <td width=264 rowspan=2 valign=top style='width:198.0pt;border-top:none;
| |
| - |
| |
| - | border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><i style='mso-bidi-font-style:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>*These constructs were used in
| |
| - | preliminary research only.<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:8;mso-yfti-lastrow:yes;height:7.8pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:7.8pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'><span style='font-size:10.0pt;font-family:
| |
| - |
| |
| - | Arial'>I0500-K152005*<o:p></o:p></span></i></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.8pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>6.62<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.8pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>4.43<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1050" type="#_x0000_t75"
| |
| - | style='width:431.25pt;height:339.75pt'>
| |
| - | <v:imagedata src="Background.files/image024.png" o:title="digestion product"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=575 height=453
| |
| - |
| |
| - | src="Background.files/image025.jpg" v:shapes="_x0000_i1050"><![endif]></p>
| |
| - |
| |
| - | <b><i style='mso-bidi-font-style:normal'><span style='font-size:16.0pt;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:SimSun;mso-font-kerning:16.0pt;
| |
| - |
| |
| - | mso-ansi-language:EN-US;mso-fareast-language:ZH-CN;mso-bidi-language:AR-SA'><br
| |
| - |
| |
| - | clear=all style='page-break-before:always'>
| |
| - | </span></i></b>
| |
| - |
| |
| - | <h1 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'>TESTING AND CHARACTERISATION</h1>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><i style='mso-bidi-font-style:normal'><u>In vivo</u></i><u> expression of
| |
| - | our devices in <i style='mso-bidi-font-style:normal'>E.coli</i> MG1655<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'>Our preliminary research indicated that carotenoids
| |
| - | production in E.coli TOP10 was very weak. We decided to use another strain, <i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'>E.coli </i>MG1655, for <i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'>in vivo </i>expression of our devices. The amount of carotenoids
| |
| - | produced became visibly higher. Generally, the colour became visible after
| |
| - | incubation for about 12 hours.<o:p></o:p></span></p>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>Lycopene-producing device (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1051" type="#_x0000_t75" style='width:234pt;height:189.75pt'>
| |
| - | <v:imagedata src="Background.files/image026.jpg" o:title="SDC12468" gain="86232f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=312 height=253
| |
| - |
| |
| - | src="Background.files/image027.jpg" v:shapes="_x0000_i1051"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.45pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1052" type="#_x0000_t75"
| |
| - | style='width:177.75pt;height:192.75pt' wrapcoords="-99 0 -99 21509 21600 21509 21600 0 -99 0">
| |
| - | <v:imagedata src="Background.files/image028.jpg" o:title="SDC12427"
| |
| - | croptop="20533f" cropbottom="8972f" cropleft="21062f" cropright="21212f"
| |
| - | gain="69719f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=237 height=257
| |
| - |
| |
| - | src="Background.files/image029.jpg" v:shapes="_x0000_i1052"><![endif]><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><i style='mso-bidi-font-style:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>E.coli </span></i><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>MG1655 transformed with K274110
| |
| - | and grown on agar plate overnight.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.45pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Cell pellet of <i>E.coli</i> MG1655
| |
| - | transformed with K274110 (from 20mL LB culture at <st1:chmetcnv UnitName="ﾰC"
| |
| - |
| |
| - | SourceValue="37" HasSpace="False" Negative="False" NumberType="1" TCSC="0"
| |
| - |
| |
| - | w:st="on">37°C</st1:chmetcnv> for 24 hours).</span><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>β -carotene-producing device
| |
| - | (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1053" type="#_x0000_t75" style='width:234pt;height:201pt'>
| |
| - | <v:imagedata src="Background.files/image030.jpg" o:title="SDC12481"
| |
| - | cropbottom="18095f" cropleft="25986f" gain="86232f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=312 height=268
| |
| - |
| |
| - | src="Background.files/image031.jpg" v:shapes="_x0000_i1053"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.7pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1054" type="#_x0000_t75" style='width:182.25pt;height:192.75pt'>
| |
| - | <v:imagedata src="Background.files/image032.jpg" o:title="SDC12432"
| |
| - | croptop="20071f" cropbottom="8971f" cropleft="21933f" cropright="17787f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=243 height=257
| |
| - |
| |
| - | src="Background.files/image033.jpg" v:shapes="_x0000_i1054"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><i style='mso-bidi-font-style:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>E.coli </span></i><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>MG1655 transformed with K274210
| |
| - | and grown on agar plate overnight.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.7pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Cell pellet of <i>E.coli</i> MG1655
| |
| - | transformed with K274210 (from 20mL LB culture at <st1:chmetcnv TCSC="0"
| |
| - |
| |
| - | NumberType="1" Negative="False" HasSpace="False" SourceValue="37"
| |
| - |
| |
| - | UnitName="ﾰC" w:st="on">37°C</st1:chmetcnv> for 24 hours).</span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <h3><u>Quantitative measurement of pigment production<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>In
| |
| - | addition to visual inspection of coloured colonies, we hope to quantitatively
| |
| - | measure the amount of lycopene or <span style='mso-bidi-font-weight:bold'>β-</span>carotene
| |
| - | produced by our devices. We grew transformed <i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'>E.coli</i> MG<st1:chmetcnv TCSC="0" NumberType="1" Negative="False"
| |
| - |
| |
| - | HasSpace="True" SourceValue="1655" UnitName="in" w:st="on">1655 in</st1:chmetcnv>
| |
| - | LB culture at </span><st1:chmetcnv TCSC="0" NumberType="1" Negative="False"
| |
| - |
| |
| - | HasSpace="False" SourceValue="37" UnitName="ﾰC" w:st="on"><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>37°C</span></st1:chmetcnv><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-bidi-font-family:"Times New Roman"'> for
| |
| - | 24 hours and collected cell pellet by centrifugation. Acetone was added to the
| |
| - | cell pellet and warmed at <st1:chmetcnv TCSC="0" NumberType="1" Negative="False"
| |
| - |
| |
| - | HasSpace="False" SourceValue="50" UnitName="ﾰC" w:st="on">50°C</st1:chmetcnv>
| |
| - | for 10 minutes, allowing carotenoids (</span><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial'>lycopene or <span style='mso-bidi-font-weight:
| |
| - |
| |
| - | bold'>β-</span>carotene) in the cells to dissolve. The acetone extracts
| |
| - | were then put into MicroPlate Reader for a full photospectrum scan (wavelengths
| |
| - | between 300nm and 800nm). The absorbance curves showed peaks characteristics of
| |
| - | the respective carotenoids: at wavelength 474nm for lycopene, and 456nm for </span><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene. In the case of β-carotene,
| |
| - | we used pure β-carotene of known amount as the standard reference and
| |
| - | included it in the graph below.</span><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'><o:p></o:p></span></p>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>Lycopene-producing device (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:307.85pt'>
| |
| - | <td width=331 valign=top style='width:248.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:307.85pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.5pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1055" type="#_x0000_t75" style='width:237.75pt;height:212.25pt'>
| |
| - | <v:imagedata src="Background.files/image034.png" o:title="lycopene_graph"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=317 height=283
| |
| - |
| |
| - | src="Background.files/image035.jpg" v:shapes="_x0000_i1055"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>Lycopene
| |
| - | in cells (pellet from 20mL LB culture at <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="37" UnitName="ﾰC"
| |
| - |
| |
| - | w:st="on">37°C</st1:chmetcnv> for 24h) was extracted with 300μL acetone.
| |
| - | For each reading, 100μL acetone exextract was diluted with 100μL
| |
| - | water and loaded on to MicroPlate Reader for a full spectrum. Each graph
| |
| - | shows the average of two readings. Value normalised with blank (50% acetone)
| |
| - | and OD<sub>600</sub>.</span><span style='font-size:10.5pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=259 valign=top style='width:194.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:307.85pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1056" type="#_x0000_t75"
| |
| - | style='width:178.5pt;height:141pt;mso-wrap-distance-left:0;
| |
| - | mso-wrap-distance-right:0;mso-position-horizontal:center' filled="t">
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image036.emz" o:title="" croptop="888f"
| |
| - | cropbottom="888f" cropleft="692f" cropright="8341f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=238 height=188
| |
| - |
| |
| - | src="Background.files/image037.gif" v:shapes="_x0000_i1056"><![endif]><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1057" type="#_x0000_t75" style='width:187.5pt;height:166.5pt'>
| |
| - | <v:imagedata src="Background.files/image038.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=250 height=222
| |
| - |
| |
| - | src="Background.files/image039.gif" v:shapes="_x0000_i1057"><![endif]><span
| |
| - |
| |
| - | style='mso-bidi-font-size:10.5pt'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>β -carotene-producing device
| |
| - | (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:263.9pt'>
| |
| - | <td width=331 valign=top style='width:248.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:263.9pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1058" type="#_x0000_t75" style='width:237.75pt;height:189pt'>
| |
| - | <v:imagedata src="Background.files/image040.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=317 height=252
| |
| - |
| |
| - | src="Background.files/image041.gif" v:shapes="_x0000_i1058"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.5pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:7.1pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span class=GramE><span style='font-size:10.0pt;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene</span></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'> in cells (pellet from 20mL LB culture
| |
| - | at <st1:chmetcnv TCSC="0" NumberType="1" Negative="False" HasSpace="False"
| |
| - |
| |
| - | SourceValue="37" UnitName="ﾰC" w:st="on">37°C</st1:chmetcnv> for 24h)
| |
| - | was extracted with 500μL acetone. 100μL acetone extract was diluted
| |
| - | with 100μL water and loaded on to MicroPlate Reader for a full spectrum
| |
| - | scan. Value normalised with blank (50% acetone) and OD<sub>600</sub>. Pure
| |
| - | carotene (from Invitrogen) was added as standard. Each graph shows average of
| |
| - | three readings.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=259 valign=top style='width:194.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:263.9pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1059" type="#_x0000_t75"
| |
| - | style='width:183.75pt;height:149.25pt;mso-wrap-distance-left:0;
| |
| - | mso-wrap-distance-right:0' o:allowoverlap="f" filled="t">
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image042.emz" o:title="" cropbottom="1187f"
| |
| - | cropright="15343f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=245 height=199
| |
| - |
| |
| - | src="Background.files/image043.gif" v:shapes="_x0000_i1059"><![endif]><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1060" type="#_x0000_t75" style='width:183.75pt;height:136.5pt;
| |
| - | mso-wrap-distance-left:0;mso-wrap-distance-right:0' wrapcoords="-62 0 -62 21502 21600 21502 21600 0 -62 0"
| |
| - | filled="t">
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image044.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=245 height=182
| |
| - |
| |
| - | src="Background.files/image045.gif" v:shapes="_x0000_i1060"><![endif]></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p style='text-align:justify;text-justify:inter-ideograph;line-height:150%'><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial'>To investigate the
| |
| - | induction of pigment production, we tested our device K<st1:chmetcnv TCSC="0"
| |
| - |
| |
| - | NumberType="1" Negative="False" HasSpace="True" SourceValue="274220"
| |
| - |
| |
| - | UnitName="in" w:st="on">274220 in</st1:chmetcnv> <i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'>E.coli</i> MG1655 and <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv>
| |
| - | arabinose was added to LB culture. Cell culture that was induced with arabinose
| |
| - | showed greater </span><span style='font-size:11.0pt;line-height:150%;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene
| |
| - | production than cell culture without arabinose, indicating the feasibility of
| |
| - | inducing pigment production. Using known amount of pure β-carotene to
| |
| - | construct a standard curve, we were able to estimate the amount of β-carotene
| |
| - | produced per unit volume of cell culture by our device.<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:303.15pt'>
| |
| - | <td width=307 valign=top style='width:230.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:303.15pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1061" type="#_x0000_t75" style='width:3in;height:169.5pt'>
| |
| - | <v:imagedata src="Background.files/image046.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=288 height=226
| |
| - |
| |
| - | src="Background.files/image047.gif" v:shapes="_x0000_i1061"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.5pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-right:6.7pt'><span style='font-size:10.0pt;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene
| |
| - | in cells (pellet from 20mL LB culture at <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="37" UnitName="ﾰC"
| |
| - |
| |
| - | w:st="on">37°C</st1:chmetcnv> for 24h; <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv>
| |
| - | arabinose was used for induction) was extracted with 500μL acetone.
| |
| - | 100μL acetone extract was diluted with 100μL water and loaded on to
| |
| - | MicroPlate Reader for a full spectrum scan. Value normalised with blank (50%
| |
| - | acetone) and OD<sub>600</sub>. Pure carotene (Invitrogen) was added as
| |
| - | standard. Each graph shows average of three readings.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=283 valign=top style='width:212.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:303.15pt'>
| |
| - | <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 align=left
| |
| - |
| |
| - | style='border-collapse:collapse;mso-table-layout-alt:fixed;mso-table-overlap:
| |
| - |
| |
| - | never;mso-table-lspace:9.0pt;margin-left:6.75pt;mso-table-rspace:9.0pt;
| |
| - |
| |
| - | margin-right:6.75pt;mso-table-anchor-vertical:paragraph;mso-table-anchor-horizontal:
| |
| - |
| |
| - | margin;mso-table-left:left;mso-table-top:162.45pt;mso-padding-alt:2.75pt 2.75pt 2.75pt 2.75pt'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:24.0pt'>
| |
| - | <td width=72 valign=top style='width:53.95pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-right:none;mso-border-top-alt:solid black .1pt;mso-border-left-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-bottom-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Samples<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=48 valign=top style='width:36.0pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-right:none;mso-border-top-alt:solid black .1pt;mso-border-left-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-bottom-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Abs. (unit)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=72 valign=top style='width:54.0pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-right:none;mso-border-top-alt:solid black .1pt;mso-border-left-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-bottom-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Carotene (μg /100μL
| |
| - | Acetone extract)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=85 valign=top style='width:63.5pt;border:solid black 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Total mass of carotene (μg
| |
| - | /mL-culture)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;height:33.7pt'>
| |
| - | <td width=72 valign=top style='width:53.95pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:33.7pt'>
| |
| - | <p class=TabellenInhalt><span lang=DE style='font-size:9.0pt;font-family:
| |
| - |
| |
| - | Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman";
| |
| - |
| |
| - | mso-ansi-language:DE'>K<st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="True" SourceValue="274210" UnitName="in" w:st="on">274210
| |
| - | in</st1:chmetcnv> <i>E.coli</i> MG1655<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=48 valign=top style='width:36.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:33.7pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>0.4<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=72 valign=top style='width:54.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:33.7pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>6<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=85 valign=top style='width:63.5pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-right-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:33.7pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>6*5/20=1.5<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:45.45pt'>
| |
| - | <td width=72 valign=top style='width:53.95pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:45.45pt'>
| |
| - | <p class=TabellenInhalt><span style='font-size:9.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>K274220,
| |
| - | with <st1:chmetcnv TCSC="0" NumberType="1" Negative="False" HasSpace="False"
| |
| - |
| |
| - | SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv> ara. in <i>E.coli</i>
| |
| - | MG1655<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=48 valign=top style='width:36.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:45.45pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>0.3<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=72 valign=top style='width:54.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:45.45pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>5<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=85 valign=top style='width:63.5pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-right-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:45.45pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>5*5/20=1.25<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1062" type="#_x0000_t75"
| |
| - | style='width:219.75pt;height:149.25pt;mso-wrap-distance-left:0;
| |
| - | mso-wrap-distance-right:0'>
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image048.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=293 height=199
| |
| - |
| |
| - | src="Background.files/image049.gif" v:shapes="_x0000_i1062"><![endif]></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <h3><u>Discussion of results<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>Despite
| |
| - | relatively poor performance in <i style='mso-bidi-font-style:normal'>E.coli</i>
| |
| - | TOP10, our pigment-producing devices gave encouraging results in <i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'>E.coli</i> MG1655. Simply by overnight
| |
| - | incubation, the colonies were able to show visible colour changes. Since the
| |
| - | precursors were already present in normal <i style='mso-bidi-font-style:normal'>E.coli</i>,
| |
| - | no additional growth medium or special treatment was needed. This paves the way
| |
| - | to developing a signalling system that is quick to produce visual outputs.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>The
| |
| - | acetone extraction of carotenoid provides a low-cost method for quantitative
| |
| - | measurement. In the above experiments, the results of photo-spectrometry were
| |
| - | consistent with characteristic absorbance wavelengths of lycopene (around
| |
| - | 475nm) and </span><span style='font-size:11.0pt;line-height:150%;font-family:
| |
| - |
| |
| - | Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene
| |
| - | (around 450nm), as noted in literature. In the case of β-carotene, the
| |
| - | absorbance profile of our β-carotene-producing device (K274210) matched
| |
| - | that of pure carotene standards, confirming that our device was functional in
| |
| - | the cells.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>When we
| |
| - | overlay the graphs of our two devices (constitutive), we obtained the following
| |
| - | result:<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:186.4pt'>
| |
| - | <td width=379 valign=top style='width:284.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:186.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;text-align:justify;text-justify:inter-ideograph;
| |
| - |
| |
| - | line-height:150%'><span style='font-size:11.0pt;line-height:150%;font-family:
| |
| - |
| |
| - | Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1063" type="#_x0000_t75" style='width:273.75pt;height:189.75pt'>
| |
| - | <v:imagedata src="Background.files/image050.gif" o:title="234"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=365 height=253
| |
| - |
| |
| - | src="Background.files/image050.gif" v:shapes="_x0000_i1063"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=211 valign=top style='width:158.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:186.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>red: K274110/lycopene;<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>orange: K274210/β-carotene;<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>black: pure β-carotene;<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>blue: untransformed <i style='mso-bidi-font-style:normal'>E.coli</i>
| |
| - | MG1655 (negative control)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>Lycopene
| |
| - | and β-carotene showed characteristic absorbance profiles, with different
| |
| - | maximum absorbance wavelengths. It was tempted to think that by converting
| |
| - | lycopene to β-carotene (e.g. by expressing CrtY gene), we would be able to
| |
| - | see a “shift” in the peaks of maximum absorbance. However, in the bar charts
| |
| - | above it was noted that lycopene and β-carotene had significant absorbance
| |
| - | at both 456nm and 474nm, albeit with different ratios (i.e. lycopene absorbed
| |
| - | more at 474nm; β-carotene absorbed more at 456nm). Therefore, measurement
| |
| - | at a single wavelength was insufficient to determine the identity of the
| |
| - | carotenoid; we might need to use “456nm:474nm” ratio instead. It would be
| |
| - | interesting to see the effect of mixing different amounts of lycopene and β-carotene,
| |
| - | and whether the “456nm:474nm” ratio can indicate the relative percentage of
| |
| - | lycopene and β-carotene. In the current project, we were unable to obtain
| |
| - | pure lycopene due to high cost of purchase.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>In terms
| |
| - | of inducing β-carotene production, <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv>
| |
| - | arabinose, acting on Pbad promoter, was able to produce a level of synthesis
| |
| - | close to that under constitutive promoter. In the non-induced population, there
| |
| - | was probably a small degree of “leakiness” and basal level of gene expression
| |
| - | (leading to some β-carotene synthesis). In future experiments, it might be
| |
| - | helpful to move the pigment-producing devices to low copy plasmids and reduce
| |
| - | the effect of such basal expression.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>One
| |
| - | possible design using our devices is to construct a system where lycopene is
| |
| - | constitutively produced and CrtY, which converts red lycopene to orange β-carotene,
| |
| - | is controlled by external stimulus. Hence, the red cells will turn orange upon
| |
| - | sensing the specific external stimulus, and this change in colour serves as the
| |
| - | signal output that can be detected visually and measured quantitatively at low
| |
| - | cost.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'><o:p> </o:p></span></p>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | </body>
| |
| - |
| |
| - | </html> <html xmlns:v="urn:schemas-microsoft-com:vml"
| |
| - |
| |
| - | xmlns:o="urn:schemas-microsoft-com:office:office"
| |
| - |
| |
| - | xmlns:w="urn:schemas-microsoft-com:office:word"
| |
| - |
| |
| - | xmlns:st1="urn:schemas-microsoft-com:office:smarttags"
| |
| - |
| |
| - | xmlns="http://www.w3.org/TR/REC-html40">
| |
| - |
| |
| - | <head>
| |
| - | <meta http-equiv=Content-Type content="text/html; charset=windows-1252">
| |
| - | <meta name=ProgId content=Word.Document>
| |
| - | <meta name=Generator content="Microsoft Word 11">
| |
| - | <meta name=Originator content="Microsoft Word 11">
| |
| - | <link rel=File-List href="Background.files/filelist.xml">
| |
| - | <link rel=Edit-Time-Data href="Background.files/editdata.mso">
| |
| - | <!--[if !mso]>
| |
| - | <style>
| |
| - | v\:* {behavior:url(#default#VML);}
| |
| - | o\:* {behavior:url(#default#VML);}
| |
| - | w\:* {behavior:url(#default#VML);}
| |
| - | .shape {behavior:url(#default#VML);}
| |
| - | </style>
| |
| - | <![endif]-->
| |
| - | <title>Background</title>
| |
| - | <o:SmartTagType namespaceuri="urn:schemas-microsoft-com:office:smarttags"
| |
| - |
| |
| - | name="City"/>
| |
| - | <o:SmartTagType namespaceuri="urn:schemas-microsoft-com:office:smarttags"
| |
| - |
| |
| - | name="place"/>
| |
| - | <o:SmartTagType namespaceuri="urn:schemas-microsoft-com:office:smarttags"
| |
| - |
| |
| - | name="chmetcnv"/>
| |
| - | <!--[if gte mso 9]><xml>
| |
| - | <o:DocumentProperties>
| |
| - | <o:Author>Siming Ma</o:Author>
| |
| - | <o:Template>Normal</o:Template>
| |
| - | <o:LastAuthor>Siming Ma</o:LastAuthor>
| |
| - | <o:Revision>2</o:Revision>
| |
| - | <o:TotalTime>454</o:TotalTime>
| |
| - | <o:Created>2009-10-18T01:28:00Z</o:Created>
| |
| - | <o:LastSaved>2009-10-18T01:28:00Z</o:LastSaved>
| |
| - | <o:Pages>1</o:Pages>
| |
| - | <o:Words>2084</o:Words>
| |
| - | <o:Characters>11885</o:Characters>
| |
| - | <o:Lines>99</o:Lines>
| |
| - | <o:Paragraphs>27</o:Paragraphs>
| |
| - | <o:CharactersWithSpaces>13942</o:CharactersWithSpaces>
| |
| - | <o:Version>11.9999</o:Version>
| |
| - | </o:DocumentProperties>
| |
| - | </xml><![endif]--><!--[if gte mso 9]><xml>
| |
| - | <w:WordDocument>
| |
| - | <w:GrammarState>Clean</w:GrammarState>
| |
| - | <w:PunctuationKerning/>
| |
| - | <w:ValidateAgainstSchemas/>
| |
| - | <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid>
| |
| - | <w:IgnoreMixedContent>false</w:IgnoreMixedContent>
| |
| - | <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText>
| |
| - | <w:Compatibility>
| |
| - | <w:BreakWrappedTables/>
| |
| - | <w:SnapToGridInCell/>
| |
| - | <w:WrapTextWithPunct/>
| |
| - | <w:UseAsianBreakRules/>
| |
| - | <w:DontGrowAutofit/>
| |
| - | <w:UseFELayout/>
| |
| - | </w:Compatibility>
| |
| - | <w:BrowserLevel>MicrosoftInternetExplorer4</w:BrowserLevel>
| |
| - | </w:WordDocument>
| |
| - | </xml><![endif]--><!--[if gte mso 9]><xml>
| |
| - | <w:LatentStyles DefLockedState="false" LatentStyleCount="156">
| |
| - | </w:LatentStyles>
| |
| - | </xml><![endif]--><!--[if !mso]><object
| |
| - | classid="clsid:38481807-CA0E-42D2-BF39-B33AF135CC4D" id=ieooui></object>
| |
| - | <style>
| |
| - | st1\:*{behavior:url(#ieooui) }
| |
| - | </style>
| |
| - | <![endif]-->
| |
| - | <style>
| |
| - | <!--
| |
| - | /* Font Definitions */
| |
| - | @font-face
| |
| - | {font-family:Wingdings;
| |
| - | panose-1:5 0 0 0 0 0 0 0 0 0;
| |
| - | mso-font-charset:2;
| |
| - | mso-generic-font-family:auto;
| |
| - | mso-font-pitch:variable;
| |
| - | mso-font-signature:0 268435456 0 0 -2147483648 0;}
| |
| - | @font-face
| |
| - | {font-family:SimSun;
| |
| - | panose-1:2 1 6 0 3 1 1 1 1 1;
| |
| - | mso-font-alt:\5B8B\4F53;
| |
| - | mso-font-charset:134;
| |
| - | mso-generic-font-family:auto;
| |
| - | mso-font-pitch:variable;
| |
| - | mso-font-signature:3 135135232 16 0 262145 0;}
| |
| - | @font-face
| |
| - | {font-family:"Arial Black";
| |
| - | panose-1:2 11 10 4 2 1 2 2 2 4;
| |
| - | mso-font-charset:0;
| |
| - | mso-generic-font-family:swiss;
| |
| - | mso-font-pitch:variable;
| |
| - | mso-font-signature:647 0 0 0 159 0;}
| |
| - | @font-face
| |
| - | {font-family:\5B8B\4F53;
| |
| - | mso-font-alt:"Times New Roman";
| |
| - | mso-font-charset:0;
| |
| - | mso-generic-font-family:auto;
| |
| - | mso-font-pitch:variable;
| |
| - | mso-font-signature:0 0 0 0 0 0;}
| |
| - | @font-face
| |
| - | {font-family:"\@SimSun";
| |
| - | panose-1:2 1 6 0 3 1 1 1 1 1;
| |
| - | mso-font-charset:134;
| |
| - | mso-generic-font-family:auto;
| |
| - | mso-font-pitch:variable;
| |
| - | mso-font-signature:3 135135232 16 0 262145 0;}
| |
| - | /* Style Definitions */
| |
| - | p.MsoNormal, li.MsoNormal, div.MsoNormal
| |
| - | {mso-style-parent:"";
| |
| - | margin:0cm;
| |
| - | margin-bottom:.0001pt;
| |
| - | mso-pagination:widow-orphan;
| |
| - | font-size:12.0pt;
| |
| - | font-family:"Times New Roman";
| |
| - | mso-fareast-font-family:SimSun;}
| |
| - | h1
| |
| - | {mso-style-next:\6B63\6587;
| |
| - | margin-top:12.0pt;
| |
| - | margin-right:0cm;
| |
| - | margin-bottom:3.0pt;
| |
| - | margin-left:0cm;
| |
| - | mso-pagination:widow-orphan;
| |
| - | page-break-after:avoid;
| |
| - | mso-outline-level:1;
| |
| - | font-size:16.0pt;
| |
| - | font-family:Arial;
| |
| - | mso-font-kerning:16.0pt;
| |
| - | font-weight:bold;}
| |
| - | h2
| |
| - | {mso-style-next:\6B63\6587;
| |
| - | margin-top:12.0pt;
| |
| - | margin-right:0cm;
| |
| - | margin-bottom:3.0pt;
| |
| - | margin-left:0cm;
| |
| - | mso-pagination:widow-orphan;
| |
| - | page-break-after:avoid;
| |
| - | mso-outline-level:2;
| |
| - | font-size:14.0pt;
| |
| - | font-family:Arial;
| |
| - | font-weight:bold;
| |
| - | font-style:italic;}
| |
| - | h3
| |
| - | {mso-style-next:\6B63\6587;
| |
| - | margin-top:12.0pt;
| |
| - | margin-right:0cm;
| |
| - | margin-bottom:3.0pt;
| |
| - | margin-left:0cm;
| |
| - | mso-pagination:widow-orphan;
| |
| - | page-break-after:avoid;
| |
| - | mso-outline-level:3;
| |
| - | font-size:13.0pt;
| |
| - | font-family:Arial;
| |
| - | font-weight:bold;}
| |
| - | a:link, span.MsoHyperlink
| |
| - | {color:blue;
| |
| - | text-decoration:underline;
| |
| - | text-underline:single;}
| |
| - | a:visited, span.MsoHyperlinkFollowed
| |
| - | {color:purple;
| |
| - | text-decoration:underline;
| |
| - | text-underline:single;}
| |
| - | p
| |
| - | {mso-margin-top-alt:auto;
| |
| - | margin-right:0cm;
| |
| - | mso-margin-bottom-alt:auto;
| |
| - | margin-left:0cm;
| |
| - | mso-pagination:widow-orphan;
| |
| - | font-size:12.0pt;
| |
| - | font-family:"Times New Roman";
| |
| - | mso-fareast-font-family:SimSun;}
| |
| - | span.mw-headline
| |
| - | {mso-style-name:mw-headline;}
| |
| - | span.apple-converted-space
| |
| - | {mso-style-name:apple-converted-space;}
| |
| - | span.apple-style-span
| |
| - | {mso-style-name:apple-style-span;}
| |
| - | p.TabellenInhalt, li.TabellenInhalt, div.TabellenInhalt
| |
| - | {mso-style-name:"Tabellen Inhalt";
| |
| - | margin:0cm;
| |
| - | margin-bottom:.0001pt;
| |
| - | mso-pagination:no-line-numbers;
| |
| - | mso-hyphenate:none;
| |
| - | font-size:12.0pt;
| |
| - | font-family:"Times New Roman";
| |
| - | mso-fareast-font-family:\5B8B\4F53;
| |
| - | mso-font-kerning:.5pt;
| |
| - | mso-fareast-language:#00FF;}
| |
| - | span.GramE
| |
| - | {mso-style-name:"";
| |
| - | mso-gram-e:yes;}
| |
| - | @page Section1
| |
| - | {size:612.0pt 792.0pt;
| |
| - | margin:9.0pt 90.0pt 18.0pt 90.0pt;
| |
| - | mso-header-margin:35.4pt;
| |
| - | mso-footer-margin:35.4pt;
| |
| - | mso-paper-source:0;}
| |
| - | div.Section1
| |
| - | {page:Section1;}
| |
| - | /* List Definitions */
| |
| - | @list l0
| |
| - | {mso-list-id:120536249;
| |
| - | mso-list-type:hybrid;
| |
| - | mso-list-template-ids:2126427888 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| |
| - | @list l0:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0B7;
| |
| - | mso-level-tab-stop:36.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | text-indent:-18.0pt;
| |
| - | font-family:Symbol;}
| |
| - | @list l1
| |
| - | {mso-list-id:329873205;
| |
| - | mso-list-type:hybrid;
| |
| - | mso-list-template-ids:422478762 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| |
| - | @list l1:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0B7;
| |
| - | mso-level-tab-stop:36.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | text-indent:-18.0pt;
| |
| - | font-family:Symbol;}
| |
| - | @list l2
| |
| - | {mso-list-id:449712540;
| |
| - | mso-list-template-ids:249083244;}
| |
| - | @list l2:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0A7;
| |
| - | mso-level-tab-stop:36.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | text-indent:-18.0pt;
| |
| - | mso-ansi-font-size:10.0pt;
| |
| - | font-family:Wingdings;}
| |
| - | @list l3
| |
| - | {mso-list-id:632642131;
| |
| - | mso-list-template-ids:-220050230;}
| |
| - | @list l3:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0A7;
| |
| - | mso-level-tab-stop:36.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | text-indent:-18.0pt;
| |
| - | mso-ansi-font-size:10.0pt;
| |
| - | font-family:Wingdings;}
| |
| - | @list l4
| |
| - | {mso-list-id:1088690536;
| |
| - | mso-list-type:hybrid;
| |
| - | mso-list-template-ids:-465797406 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| |
| - | @list l4:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0B7;
| |
| - | mso-level-tab-stop:72.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | margin-left:72.0pt;
| |
| - | text-indent:-18.0pt;
| |
| - | font-family:Symbol;}
| |
| - | @list l5
| |
| - | {mso-list-id:1133330608;
| |
| - | mso-list-type:hybrid;
| |
| - | mso-list-template-ids:1158428478 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| |
| - | @list l5:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0B7;
| |
| - | mso-level-tab-stop:36.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | text-indent:-18.0pt;
| |
| - | font-family:Symbol;}
| |
| - | @list l6
| |
| - | {mso-list-id:1410152836;
| |
| - | mso-list-template-ids:-1665133950;}
| |
| - | @list l6:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0A7;
| |
| - | mso-level-tab-stop:36.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | text-indent:-18.0pt;
| |
| - | mso-ansi-font-size:10.0pt;
| |
| - | font-family:Wingdings;}
| |
| - | @list l7
| |
| - | {mso-list-id:2013410533;
| |
| - | mso-list-type:hybrid;
| |
| - | mso-list-template-ids:-248105022 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;}
| |
| - | @list l7:level1
| |
| - | {mso-level-number-format:bullet;
| |
| - | mso-level-text:\F0B7;
| |
| - | mso-level-tab-stop:18.0pt;
| |
| - | mso-level-number-position:left;
| |
| - | margin-left:18.0pt;
| |
| - | text-indent:-18.0pt;
| |
| - | font-family:Symbol;}
| |
| - | ol
| |
| - | {margin-bottom:0cm;}
| |
| - | ul
| |
| - | {margin-bottom:0cm;}
| |
| - | -->
| |
| - | </style>
| |
| - | <!--[if gte mso 10]>
| |
| - | <style>
| |
| - | /* Style Definitions */
| |
| - | table.MsoNormalTable
| |
| - | {mso-style-name:\666E\901A\8868\683C;
| |
| - | mso-tstyle-rowband-size:0;
| |
| - | mso-tstyle-colband-size:0;
| |
| - | mso-style-noshow:yes;
| |
| - | mso-style-parent:"";
| |
| - | mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
| |
| - | mso-para-margin:0cm;
| |
| - | mso-para-margin-bottom:.0001pt;
| |
| - | mso-pagination:widow-orphan;
| |
| - | font-size:10.0pt;
| |
| - | font-family:"Times New Roman";
| |
| - | mso-ansi-language:#0400;
| |
| - | mso-fareast-language:#0400;
| |
| - | mso-bidi-language:#0400;}
| |
| - | table.MsoTableGrid
| |
| - | {mso-style-name:\7F51\683C\578B;
| |
| - | mso-tstyle-rowband-size:0;
| |
| - | mso-tstyle-colband-size:0;
| |
| - | border:solid windowtext 1.0pt;
| |
| - | mso-border-alt:solid windowtext .5pt;
| |
| - | mso-padding-alt:0cm 5.4pt 0cm 5.4pt;
| |
| - | mso-border-insideh:.5pt solid windowtext;
| |
| - | mso-border-insidev:.5pt solid windowtext;
| |
| - | mso-para-margin:0cm;
| |
| - | mso-para-margin-bottom:.0001pt;
| |
| - | mso-pagination:widow-orphan;
| |
| - | font-size:10.0pt;
| |
| - | font-family:"Times New Roman";
| |
| - | mso-ansi-language:#0400;
| |
| - | mso-fareast-language:#0400;
| |
| - | mso-bidi-language:#0400;}
| |
| - | </style>
| |
| - | <![endif]--><!--[if gte mso 9]><xml>
| |
| - | <o:shapedefaults v:ext="edit" spidmax="2050"/>
| |
| - | </xml><![endif]--><!--[if gte mso 9]><xml>
| |
| - | <o:shapelayout v:ext="edit">
| |
| - | <o:idmap v:ext="edit" data="1"/>
| |
| - | </o:shapelayout></xml><![endif]-->
| |
| - | </head>
| |
| - |
| |
| - | <body lang=EN-US link=blue vlink=purple style='tab-interval:36.0pt'>
| |
| - |
| |
| - | <div class=Section1>
| |
| - |
| |
| - | <h1 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'>BACKGROUND</h1>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;text-align:justify;text-justify:inter-ideograph;line-height:150%'><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-weight:normal;mso-bidi-font-weight:
| |
| - |
| |
| - | bold'>Carotenoids are organic pigments naturally present in plants, algae and
| |
| - | some bacteria. There are more than 600 carotenoids, which can be categorized
| |
| - | into xanthophylls (hydrocarbons containing oxygen) and carotenes (hydrocarbons
| |
| - | containing no oxygen). Carotenoids perform a range of functions, including
| |
| - | light energy absorption, protection against photo-damage, acting as
| |
| - | antioxidants, and as precursor to other organic compounds. In human, for
| |
| - | example, beta-carotene is the precursor to vitamin A.<o:p></o:p></span></h3>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><u>Biochemical pathway of carotenoid synthesis<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;text-align:justify;text-justify:inter-ideograph;line-height:150%'><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial'>The common starting
| |
| - | point for carotenoid synthesis is<span class=apple-converted-space> </span><span
| |
| - |
| |
| - | style='mso-bidi-font-weight:bold'>farnesyl pyrophosphate (FPP)</span>, which
| |
| - | derives from two precursors, isopentenyl pyrophosphate (IPP) and dimethylallyl
| |
| - | pyrophosphate (DMAPP). In general, there are two pathways for synthesising IPP
| |
| - | and DMAPP: the Mevalonate Pathway (starting with acetyl CoA) and the
| |
| - | Non-mevalonate Pathway (starting with pyruvate and glyceradehyde-3-phosphate).
| |
| - | While the Mevalonate Pathway is present in all higher eukaryotes, the<span
| |
| - |
| |
| - | class=apple-converted-space> </span><span style='mso-bidi-font-weight:
| |
| - |
| |
| - | bold'>Non-mevalonate Pathway</span><span class=apple-converted-space> </span>is
| |
| - | present in<span class=apple-converted-space> </span><i>E. coli</i>.<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
| |
| - | <td width=307 valign=top style='width:230.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center;background:#F9F9F9'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Cam09_Mevalonate_pathway.png"
| |
| - |
| |
| - | title="Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)"><!--[if gte vml 1]><v:shapetype
| |
| - | id="_x0000_t75" coordsize="21600,21600" o:spt="75" o:preferrelative="t"
| |
| - | path="m@4@5l@4@11@9@11@9@5xe" filled="f" stroked="f">
| |
| - | <v:stroke joinstyle="miter"/>
| |
| - | <v:formulas>
| |
| - | <v:f eqn="if lineDrawn pixelLineWidth 0"/>
| |
| - | <v:f eqn="sum @0 1 0"/>
| |
| - | <v:f eqn="sum 0 0 @1"/>
| |
| - | <v:f eqn="prod @2 1 2"/>
| |
| - | <v:f eqn="prod @3 21600 pixelWidth"/>
| |
| - | <v:f eqn="prod @3 21600 pixelHeight"/>
| |
| - | <v:f eqn="sum @0 0 1"/>
| |
| - | <v:f eqn="prod @6 1 2"/>
| |
| - | <v:f eqn="prod @7 21600 pixelWidth"/>
| |
| - | <v:f eqn="sum @8 21600 0"/>
| |
| - | <v:f eqn="prod @7 21600 pixelHeight"/>
| |
| - | <v:f eqn="sum @10 21600 0"/>
| |
| - | </v:formulas>
| |
| - | <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>
| |
| - | <o:lock v:ext="edit" aspectratio="t"/>
| |
| - | </v:shapetype><v:shape id="_x0000_i1025" type="#_x0000_t75" alt="Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)"
| |
| - | title=""Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)""
| |
| - | style='width:214.5pt;height:132.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image001.png" o:href="https://static.igem.org/mediawiki/2009/thumb/d/d1/Cam09_Mevalonate_pathway.png/334px-Cam09_Mevalonate_pathway.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=286 height=177
| |
| - |
| |
| - | src="Background.files/image002.gif"
| |
| - |
| |
| - | alt="Mevalonate Pathway (image adopted from Wikipedia, http://en.wikipedia.org/wiki/Mevalonate_pathway)"
| |
| - |
| |
| - | v:shapes="_x0000_i1025"><![endif]></a><o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><b><span style='font-size:8.5pt;font-family:Arial'>Mevalonate Pathway</span></b><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:8.5pt;font-family:Arial'> </span></span><span
| |
| - |
| |
| - | style='font-size:8.5pt;font-family:Arial'>(image adopted from Wikipedia, <a
| |
| - |
| |
| - | href="http://en.wikipedia.org/wiki/Mevalonate_pathway"
| |
| - |
| |
| - | title="http://en.wikipedia.org/wiki/Mevalonate_pathway"><span
| |
| - |
| |
| - | style='color:windowtext;text-decoration:none;text-underline:none'>http://en.wikipedia.org/wiki/Mevalonate_pathway</span></a>)</span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=283 valign=top style='width:212.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:9.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Cam09_Non_mevalonate.JPG"
| |
| - |
| |
| - | title="Non-mevalonate Pathway (image adopted from Yuan et al. 2006)"><span
| |
| - |
| |
| - | style='color:windowtext;text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1026" type="#_x0000_t75" alt="Non-mevalonate Pathway (image adopted from Yuan et al. 2006)"
| |
| - | title=""Non-mevalonate Pathway (image adopted from Yuan et al. 2006)""
| |
| - | style='width:201.75pt;height:141pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image003.jpg" o:href="https://static.igem.org/mediawiki/2009/thumb/1/19/Cam09_Non_mevalonate.JPG/360px-Cam09_Non_mevalonate.JPG"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=269 height=188
| |
| - |
| |
| - | src="Background.files/image004.jpg"
| |
| - |
| |
| - | alt="Non-mevalonate Pathway (image adopted from Yuan et al. 2006)" v:shapes="_x0000_i1026"><![endif]></span></a></span><b><span
| |
| - |
| |
| - | style='font-size:8.5pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>Non-mevalonate Pathway</span></b><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:8.5pt;font-family:Arial'> </span></span><span
| |
| - |
| |
| - | style='font-size:8.5pt;font-family:Arial'>(image adopted from Yuan et al.
| |
| - | 2006)</span><span style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;text-align:justify;text-justify:inter-ideograph'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'><br>
| |
| - | </span><span style='font-size:11.0pt;font-family:Arial'>After IPP and DMAPP go
| |
| - | on to form FPP, a series of enzymatic reactions convert the colourless FPP to
| |
| - | coloured compounds: red lycopene, and orange β-carotene.<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes'>
| |
| - | <td width=590 valign=top style='width:442.8pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1027" type="#_x0000_t75" style='width:6in;height:282.75pt'>
| |
| - | <v:imagedata src="Background.files/image005.jpg" o:title="Cambridge_Carotenoid_100"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=576 height=377
| |
| - |
| |
| - | src="Background.files/image006.jpg" v:shapes="_x0000_i1027"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span class=apple-style-span><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial;mso-bidi-font-weight:bold'>By modulating the enzymes
| |
| - | involved in this conversion pathway ("CrtE", "CrtB",
| |
| - | "CrtI", and "CrtY"), we aim to produce and control
| |
| - | different pigments as signal output.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <b><span style='font-size:16.0pt;font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | SimSun;mso-font-kerning:16.0pt;mso-ansi-language:EN-US;mso-fareast-language:
| |
| - |
| |
| - | ZH-CN;mso-bidi-language:AR-SA'><br clear=all style='page-break-before:always'>
| |
| - | </span></b>
| |
| - |
| |
| - | <h1 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'>DESIGN OF DEVICES</h1>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><u>Biobricks contributed by previous iGEM teams<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>A few iGEM
| |
| - | teams of previous years (e.g. <st1:City w:st="on">Edinburgh</st1:City> 07, <st1:City
| |
| - |
| |
| - | w:st="on">Edinburgh</st1:City> 08, <st1:place w:st="on"><st1:City w:st="on"><span
| |
| - |
| |
| - | class=GramE>Guelph</span></st1:City></st1:place> 08) conducted research in
| |
| - | the carotenoid system. We are fortunate to have access to some of their
| |
| - | Biobricks in the registry:<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoNormalTable border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='mso-cellspacing:0cm;background:#3D5089;mso-padding-alt:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=67 style='width:50.55pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Registry Code<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Team<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Sequence Description<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal align=center style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:center'><b><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Size of insert<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118014<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1028"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1028"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtE)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>922bp<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118006<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1029"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1029"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtB)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>949bp<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:3'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118005<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1030"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1030"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtI)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>1498bp<o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'>2 PstI sites removed.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:4'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K118013<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Edinburgh</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1031"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1031"><![endif]></span></a><span
| |
| - |
| |
| - | class=apple-converted-space> </span>(rbs + CrtY)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>1162bp<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:5;mso-yfti-lastrow:yes'>
| |
| - | <td width=67 style='width:50.55pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>K152005<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=96 style='width:71.7pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><st1:City w:st="on"><st1:place w:st="on"><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Guelph</span></st1:place></st1:City><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'> 08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=252 style='width:189.0pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1032"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1032"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1033"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1033"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1034"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1034"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_translational_unit.png"
| |
| - |
| |
| - | title="Image:Icon translational unit.png"><span style='color:windowtext;
| |
| - |
| |
| - | text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape id="_x0000_i1035"
| |
| - | type="#_x0000_t75" alt="Image:Icon translational unit.png" title=""Image:Icon translational unit.png""
| |
| - | style='width:34.5pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image007.png" o:href="https://static.igem.org/mediawiki/2009/9/91/Icon_translational_unit.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=46 height=25
| |
| - |
| |
| - | src="Background.files/image008.gif" alt="Image:Icon translational unit.png"
| |
| - |
| |
| - | v:shapes="_x0000_i1035"><![endif]></span></a><a
| |
| - |
| |
| - | href="/Image:Icon_reporter.png" title="Image:Icon reporter.png"><span
| |
| - |
| |
| - | style='color:windowtext;text-decoration:none;text-underline:none'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1036" type="#_x0000_t75" alt="Image:Icon reporter.png" title=""Image:Icon reporter.png""
| |
| - | style='width:27.75pt;height:18.75pt' o:button="t">
| |
| - | <v:imagedata src="Background.files/image009.png" o:href="https://static.igem.org/mediawiki/2009/a/aa/Icon_reporter.png"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=37 height=25
| |
| - |
| |
| - | src="Background.files/image010.jpg" alt="Image:Icon reporter.png" v:shapes="_x0000_i1036"><![endif]></span></a><o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'>(rbs+CrtE) + (rbs+CrtB)
| |
| - | + (rbs+CrtI) + (rbs+CrtY) + (rbs+GFP)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=175 style='width:131.25pt;background:white;padding:4.5pt 4.5pt 4.5pt 4.5pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>5412bp<o:p></o:p></span></p>
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:10.0pt;font-family:Arial'>2 PstI sites in CrtI
| |
| - | removed.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span class=apple-converted-space><span style='font-size:
| |
| - |
| |
| - | 11.0pt;mso-bidi-font-size:12.0pt;line-height:150%;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>The above enzymes all come from a type of Gram negative
| |
| - | Enterobacteria, <i style='mso-bidi-font-style:normal'>Pantoea ananatis</i> (GenBank:
| |
| - | D90087.2). It is noted that analogues of these enzymes are also found in other
| |
| - | organisms (e.g. bacteria <i style='mso-bidi-font-style:normal'>Rhodobacter</i>
| |
| - | spp. and the fungus <i style='mso-bidi-font-style:normal'>Neurospora crassa</i>).<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span class=apple-converted-space><span style='font-size:
| |
| - |
| |
| - | 11.0pt;mso-bidi-font-size:12.0pt;line-height:150%;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>During our preliminary research, we have encountered a few
| |
| - | issues:<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:35.7pt;text-indent:-17.85pt;mso-list:l5 level1 lfo6;tab-stops:list 36.0pt'><![if !supportLists]><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:11.0pt;mso-bidi-font-size:
| |
| - |
| |
| - | 12.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
| |
| - |
| |
| - | Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>
| |
| - | </span></span></span></span><![endif]><span class=apple-converted-space><span
| |
| - |
| |
| - | style='font-size:11.0pt;mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>Originally, the registry annotation for K152005 indicated
| |
| - | that there was no ribosome binding site before gene CrtI (i.e.
| |
| - | “…rbs+CrtB+CrtI+rbs+CrtY…”). After clarification with Team Guelph 08, we were
| |
| - | told that it was an error in annotation. The registry entry for K152005 has
| |
| - | since been updated to include the ribosome binding site.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:35.7pt;text-indent:-17.85pt;mso-list:l5 level1 lfo6;tab-stops:list 36.0pt'><![if !supportLists]><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:11.0pt;mso-bidi-font-size:
| |
| - |
| |
| - | 12.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
| |
| - |
| |
| - | Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>
| |
| - | </span></span></span></span><![endif]><span class=apple-converted-space><span
| |
| - |
| |
| - | style='font-size:11.0pt;mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>Teams of previous years constructed various composite parts
| |
| - | containing two or more enzymes of the synthetic pathways (e.g. CrtEBI,
| |
| - | CrtEBIY), but they are not available on the 2009 Distribution Plates.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:35.7pt;text-indent:-17.85pt;mso-list:l5 level1 lfo6;tab-stops:list 36.0pt'><![if !supportLists]><span
| |
| - |
| |
| - | class=apple-converted-space><span style='font-size:11.0pt;mso-bidi-font-size:
| |
| - |
| |
| - | 12.0pt;font-family:Symbol;mso-fareast-font-family:Symbol;mso-bidi-font-family:
| |
| - |
| |
| - | Symbol'><span style='mso-list:Ignore'>·<span style='font:7.0pt "Times New Roman"'>
| |
| - | </span></span></span></span><![endif]><span class=apple-converted-space><span
| |
| - |
| |
| - | style='font-size:11.0pt;mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>We put K152005 under control of constitutive promoter
| |
| - | (R0011) and transformed the construct into E.coli TOP10 cells. However, the
| |
| - | plasmids seemed unstable (the colonies streaks on solid agar plates were
| |
| - | inconsistent in colour) and the pigment production was low. This indicated that
| |
| - | a different strain of E.coli might be needed for pigment production.<o:p></o:p></span></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><span class=apple-converted-space><span style='font-size:11.0pt;
| |
| - |
| |
| - | mso-bidi-font-size:12.0pt;font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></span></p>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><u>Create our own devices by standard assembly<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><span style='font-size:11.0pt;font-family:Arial'>Given the
| |
| - | information at hand, we decided to adopt the following general strategies:<o:p></o:p></span></p>
| |
| - |
| |
| - | <ul style='margin-top:0cm' type=disc>
| |
| - | <li class=MsoNormal style='margin-top:6.0pt;margin-bottom:6.0pt;mso-list:l0 level1 lfo7;
| |
| - |
| |
| - | tab-stops:list 36.0pt'><span style='font-size:11.0pt;font-family:Arial'>Create
| |
| - | a lycopene-producing device (with enzymes CrtE, CrtB and CrtI) by standard
| |
| - | assembly of Biobricks K118014, K118006 and K118005.<o:p></o:p></span></li>
| |
| - | <li class=MsoNormal style='margin-top:6.0pt;margin-bottom:6.0pt;mso-list:l0 level1 lfo7;
| |
| - |
| |
| - | tab-stops:list 36.0pt'><span style='font-size:11.0pt;font-family:Arial'>Create
| |
| - | a <span style='mso-bidi-font-weight:bold'>β-carotene-producing device
| |
| - | (with enzymes CrtE, CrtB, CrtI and CrtY) by standard assembly of Biobricks
| |
| - | K118014, K118006, K118005 and K118013.</span><o:p></o:p></span></li>
| |
| - | <li class=MsoNormal style='margin-top:6.0pt;margin-bottom:6.0pt;mso-list:l0 level1 lfo7;
| |
| - |
| |
| - | tab-stops:list 36.0pt'><span style='font-size:11.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-weight:bold'>Put the above two constructs under constitutive
| |
| - | promoter (R0011) and arabinose-induced promoter/Pbad (I0500) and test the
| |
| - | effect on colour production.</span><span style='font-size:11.0pt;
| |
| - |
| |
| - | font-family:Arial'><o:p></o:p></span></li>
| |
| - | </ul>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:11.0pt'>Scheme for standard assembly<o:p></o:p></span></h2>
| |
| - |
| |
| - | <div align=center>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=90 valign=top style='width:67.5pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial;mso-fareast-font-family:Arial'>Lycopene-producing
| |
| - | device<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b><span style='font-size:10.0pt;font-family:Arial'>β-carotene-producing
| |
| - | device</span></b><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial;mso-fareast-font-family:Arial'><o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Constituent
| |
| - | Biobricks (already in the registry)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1037"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropright="41936f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image012.jpg" v:shapes="_x0000_i1037"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118014)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1038"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="21948f" cropright="21948f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image013.jpg" v:shapes="_x0000_i1038"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118006)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1039"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="42249f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image014.jpg" v:shapes="_x0000_i1039"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118005)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1040"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropright="41936f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image012.jpg" v:shapes="_x0000_i1040"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118014)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1041"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="21948f" cropright="21948f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image013.jpg" v:shapes="_x0000_i1041"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118006)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1042"
| |
| - | type="#_x0000_t75" style='width:58.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"
| |
| - | cropbottom="-2252f" cropleft="42249f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=78 height=33
| |
| - |
| |
| - | src="Background.files/image014.jpg" v:shapes="_x0000_i1042"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118005)<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1043"
| |
| - | type="#_x0000_t75" style='width:59.25pt;height:24pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"
| |
| - | cropbottom="-2030f" cropleft="50400f" grayscale="t"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=79 height=32
| |
| - |
| |
| - | src="Background.files/image016.jpg" v:shapes="_x0000_i1043"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><span
| |
| - |
| |
| - | style='mso-spacerun:yes'> </span>(K118013)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Basic construct<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1044" type="#_x0000_t75"
| |
| - | style='width:135pt;height:24pt'>
| |
| - | <v:imagedata src="Background.files/image011.png" o:title="untitled"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=180 height=32
| |
| - |
| |
| - | src="Background.files/image017.jpg" v:shapes="_x0000_i1044"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial'>K274100</span></b><span style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1045" type="#_x0000_t75"
| |
| - | style='width:160.5pt;height:24pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"
| |
| - | cropbottom="-2114f" cropleft="9882f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=214 height=32
| |
| - |
| |
| - | src="Background.files/image018.jpg" v:shapes="_x0000_i1045"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial'>K274200</span></b><span style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:3'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Under constitutive promoter<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1046" type="#_x0000_t75"
| |
| - | style='width:155.25pt;height:26.25pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"
| |
| - | cropbottom="-2427f" cropright="13653f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=207 height=35
| |
| - |
| |
| - | src="Background.files/image019.jpg" v:shapes="_x0000_i1046"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial;mso-fareast-font-family:Arial'>K274110</span></b><span
| |
| - |
| |
| - | style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1047" type="#_x0000_t75"
| |
| - | style='width:170.25pt;height:23.25pt'>
| |
| - | <v:imagedata src="Background.files/image015.png" o:title="untitled2"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=227 height=31
| |
| - |
| |
| - | src="Background.files/image020.jpg" v:shapes="_x0000_i1047"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial;mso-fareast-font-family:Arial'>K274210</span></b><span
| |
| - |
| |
| - | style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:4;mso-yfti-lastrow:yes'>
| |
| - | <td width=90 style='width:67.5pt;border:solid windowtext 1.0pt;border-top:
| |
| - |
| |
| - | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Under Pbad promoter<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=229 valign=top style='width:171.9pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1048" type="#_x0000_t75"
| |
| - | style='width:157.5pt;height:24.75pt'>
| |
| - | <v:imagedata src="Background.files/image021.png" o:title="untitled3"
| |
| - | cropright="12765f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=210 height=33
| |
| - |
| |
| - | src="Background.files/image022.jpg" v:shapes="_x0000_i1048"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial;mso-fareast-font-family:Arial'>K274120</span></b><span
| |
| - |
| |
| - | style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=241 valign=top style='width:181.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial'><!--[if gte vml 1]><v:shape id="_x0000_i1049" type="#_x0000_t75"
| |
| - | style='width:170.25pt;height:23.25pt'>
| |
| - | <v:imagedata src="Background.files/image021.png" o:title="untitled3"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=227 height=31
| |
| - |
| |
| - | src="Background.files/image023.jpg" v:shapes="_x0000_i1049"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-family:Arial'>K274220</span></b><span style='font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span style='font-size:11.0pt;line-height:150%;
| |
| - |
| |
| - | font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm;line-height:150%'><span style='font-size:11.0pt;line-height:150%;
| |
| - |
| |
| - | font-family:Arial;mso-bidi-font-family:"Times New Roman"'>After constructing
| |
| - | our devices, we conducted restriction digest (cut with enzymes XbaI and PstI on
| |
| - | Biobrick prefix and suffix, respectively) to check that the sizes of the
| |
| - | inserts were correct. The results are as follows:<o:p></o:p></span></p>
| |
| - |
| |
| - | <div align=center>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:24.55pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Biobrick Number<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Size of insert (kb)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Size of backbone (kb)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:24.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>Note<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;height:7.6pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274100<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>3.37<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.6pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Digestion
| |
| - | result not shown.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2;height:8.05pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274110<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>3.42<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.05pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:3;height:17.55pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274120<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.58<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.43<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:17.55pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Sizes
| |
| - | of insert and backbone were similar so only one band appeared on gel.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:4;height:8.95pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274200<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.53<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.95pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'>Digestion
| |
| - | result not shown.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:5;height:8.1pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274210<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.59<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>2.08<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.1pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:6;height:8.15pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>K274220<o:p></o:p></span></b></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>5.74<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'>4.43<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=264 valign=top style='width:198.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:8.15pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:7;height:23.25pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'><span style='font-size:10.0pt;font-family:
| |
| - |
| |
| - | Arial'>R0011-K152005*<o:p></o:p></span></i></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>5.48<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>2.08<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | <td width=264 rowspan=2 valign=top style='width:198.0pt;border-top:none;
| |
| - |
| |
| - | border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:23.25pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><i style='mso-bidi-font-style:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>*These constructs were used in
| |
| - | preliminary research only.<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:8;mso-yfti-lastrow:yes;height:7.8pt'>
| |
| - | <td width=92 valign=top style='width:68.85pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt;height:7.8pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><b style='mso-bidi-font-weight:normal'><i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'><span style='font-size:10.0pt;font-family:
| |
| - |
| |
| - | Arial'>I0500-K152005*<o:p></o:p></span></i></b></p>
| |
| - | </td>
| |
| - | <td width=84 valign=top style='width:63.0pt;border-top:none;border-left:none;
| |
| - |
| |
| - | border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.8pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>6.62<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | <td width=108 valign=top style='width:81.0pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:7.8pt'>
| |
| - | <p class=MsoNormal align=right style='margin-top:6.0pt;margin-right:0cm;
| |
| - |
| |
| - | margin-bottom:6.0pt;margin-left:0cm;text-align:right'><i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'><span style='font-size:10.0pt;font-family:Arial'>4.43<o:p></o:p></span></i></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1050" type="#_x0000_t75"
| |
| - | style='width:431.25pt;height:339.75pt'>
| |
| - | <v:imagedata src="Background.files/image024.png" o:title="digestion product"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=575 height=453
| |
| - |
| |
| - | src="Background.files/image025.jpg" v:shapes="_x0000_i1050"><![endif]></p>
| |
| - |
| |
| - | <b><i style='mso-bidi-font-style:normal'><span style='font-size:16.0pt;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:SimSun;mso-font-kerning:16.0pt;
| |
| - |
| |
| - | mso-ansi-language:EN-US;mso-fareast-language:ZH-CN;mso-bidi-language:AR-SA'><br
| |
| - |
| |
| - | clear=all style='page-break-before:always'>
| |
| - | </span></i></b>
| |
| - |
| |
| - | <h1 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'>TESTING AND CHARACTERISATION</h1>
| |
| - |
| |
| - | <h3 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><i style='mso-bidi-font-style:normal'><u>In vivo</u></i><u> expression of
| |
| - | our devices in <i style='mso-bidi-font-style:normal'>E.coli</i> MG1655<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'>Our preliminary research indicated that carotenoids
| |
| - | production in E.coli TOP10 was very weak. We decided to use another strain, <i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'>E.coli </i>MG1655, for <i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'>in vivo </i>expression of our devices. The amount of carotenoids
| |
| - | produced became visibly higher. Generally, the colour became visible after
| |
| - | incubation for about 12 hours.<o:p></o:p></span></p>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>Lycopene-producing device (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1051" type="#_x0000_t75" style='width:234pt;height:189.75pt'>
| |
| - | <v:imagedata src="Background.files/image026.jpg" o:title="SDC12468" gain="86232f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=312 height=253
| |
| - |
| |
| - | src="Background.files/image027.jpg" v:shapes="_x0000_i1051"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.45pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1052" type="#_x0000_t75"
| |
| - | style='width:177.75pt;height:192.75pt' wrapcoords="-99 0 -99 21509 21600 21509 21600 0 -99 0">
| |
| - | <v:imagedata src="Background.files/image028.jpg" o:title="SDC12427"
| |
| - | croptop="20533f" cropbottom="8972f" cropleft="21062f" cropright="21212f"
| |
| - | gain="69719f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=237 height=257
| |
| - |
| |
| - | src="Background.files/image029.jpg" v:shapes="_x0000_i1052"><![endif]><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><i style='mso-bidi-font-style:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>E.coli </span></i><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>MG1655 transformed with K274110
| |
| - | and grown on agar plate overnight.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.45pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Cell pellet of <i>E.coli</i> MG1655
| |
| - | transformed with K274110 (from 20mL LB culture at <st1:chmetcnv UnitName="ﾰC"
| |
| - |
| |
| - | SourceValue="37" HasSpace="False" Negative="False" NumberType="1" TCSC="0"
| |
| - |
| |
| - | w:st="on">37°C</st1:chmetcnv> for 24 hours).</span><span style='font-size:
| |
| - |
| |
| - | 10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>β -carotene-producing device
| |
| - | (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1053" type="#_x0000_t75" style='width:234pt;height:201pt'>
| |
| - | <v:imagedata src="Background.files/image030.jpg" o:title="SDC12481"
| |
| - | cropbottom="18095f" cropleft="25986f" gain="86232f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=312 height=268
| |
| - |
| |
| - | src="Background.files/image031.jpg" v:shapes="_x0000_i1053"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.7pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1054" type="#_x0000_t75" style='width:182.25pt;height:192.75pt'>
| |
| - | <v:imagedata src="Background.files/image032.jpg" o:title="SDC12432"
| |
| - | croptop="20071f" cropbottom="8971f" cropleft="21933f" cropright="17787f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=243 height=257
| |
| - |
| |
| - | src="Background.files/image033.jpg" v:shapes="_x0000_i1054"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;mso-yfti-lastrow:yes'>
| |
| - | <td width=303 valign=top style='width:226.95pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><i style='mso-bidi-font-style:normal'><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>E.coli </span></i><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'>MG1655 transformed with K274210
| |
| - | and grown on agar plate overnight.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=233 valign=top style='width:174.7pt;border-top:none;border-left:
| |
| - |
| |
| - | none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Cell pellet of <i>E.coli</i> MG1655
| |
| - | transformed with K274210 (from 20mL LB culture at <st1:chmetcnv TCSC="0"
| |
| - |
| |
| - | NumberType="1" Negative="False" HasSpace="False" SourceValue="37"
| |
| - |
| |
| - | UnitName="ﾰC" w:st="on">37°C</st1:chmetcnv> for 24 hours).</span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <h3><u>Quantitative measurement of pigment production<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>In
| |
| - | addition to visual inspection of coloured colonies, we hope to quantitatively
| |
| - | measure the amount of lycopene or <span style='mso-bidi-font-weight:bold'>β-</span>carotene
| |
| - | produced by our devices. We grew transformed <i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'>E.coli</i> MG<st1:chmetcnv TCSC="0" NumberType="1" Negative="False"
| |
| - |
| |
| - | HasSpace="True" SourceValue="1655" UnitName="in" w:st="on">1655 in</st1:chmetcnv>
| |
| - | LB culture at </span><st1:chmetcnv TCSC="0" NumberType="1" Negative="False"
| |
| - |
| |
| - | HasSpace="False" SourceValue="37" UnitName="ﾰC" w:st="on"><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>37°C</span></st1:chmetcnv><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-bidi-font-family:"Times New Roman"'> for
| |
| - | 24 hours and collected cell pellet by centrifugation. Acetone was added to the
| |
| - | cell pellet and warmed at <st1:chmetcnv TCSC="0" NumberType="1" Negative="False"
| |
| - |
| |
| - | HasSpace="False" SourceValue="50" UnitName="ﾰC" w:st="on">50°C</st1:chmetcnv>
| |
| - | for 10 minutes, allowing carotenoids (</span><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial'>lycopene or <span style='mso-bidi-font-weight:
| |
| - |
| |
| - | bold'>β-</span>carotene) in the cells to dissolve. The acetone extracts
| |
| - | were then put into MicroPlate Reader for a full photospectrum scan (wavelengths
| |
| - | between 300nm and 800nm). The absorbance curves showed peaks characteristics of
| |
| - | the respective carotenoids: at wavelength 474nm for lycopene, and 456nm for </span><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial;mso-fareast-font-family:
| |
| - |
| |
| - | Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene. In the case of β-carotene,
| |
| - | we used pure β-carotene of known amount as the standard reference and
| |
| - | included it in the graph below.</span><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'><o:p></o:p></span></p>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>Lycopene-producing device (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:307.85pt'>
| |
| - | <td width=331 valign=top style='width:248.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:307.85pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.5pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1055" type="#_x0000_t75" style='width:237.75pt;height:212.25pt'>
| |
| - | <v:imagedata src="Background.files/image034.png" o:title="lycopene_graph"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=317 height=283
| |
| - |
| |
| - | src="Background.files/image035.jpg" v:shapes="_x0000_i1055"><![endif]><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>Lycopene
| |
| - | in cells (pellet from 20mL LB culture at <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="37" UnitName="ﾰC"
| |
| - |
| |
| - | w:st="on">37°C</st1:chmetcnv> for 24h) was extracted with 300μL acetone.
| |
| - | For each reading, 100μL acetone exextract was diluted with 100μL
| |
| - | water and loaded on to MicroPlate Reader for a full spectrum. Each graph
| |
| - | shows the average of two readings. Value normalised with blank (50% acetone)
| |
| - | and OD<sub>600</sub>.</span><span style='font-size:10.5pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=259 valign=top style='width:194.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:307.85pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1056" type="#_x0000_t75"
| |
| - | style='width:178.5pt;height:141pt;mso-wrap-distance-left:0;
| |
| - | mso-wrap-distance-right:0;mso-position-horizontal:center' filled="t">
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image036.emz" o:title="" croptop="888f"
| |
| - | cropbottom="888f" cropleft="692f" cropright="8341f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=238 height=188
| |
| - |
| |
| - | src="Background.files/image037.gif" v:shapes="_x0000_i1056"><![endif]><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1057" type="#_x0000_t75" style='width:187.5pt;height:166.5pt'>
| |
| - | <v:imagedata src="Background.files/image038.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=250 height=222
| |
| - |
| |
| - | src="Background.files/image039.gif" v:shapes="_x0000_i1057"><![endif]><span
| |
| - |
| |
| - | style='mso-bidi-font-size:10.5pt'><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <h2 style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;margin-left:
| |
| - |
| |
| - | 0cm'><span style='font-size:12.0pt'>β -carotene-producing device
| |
| - | (constitutive)<o:p></o:p></span></h2>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:263.9pt'>
| |
| - | <td width=331 valign=top style='width:248.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:263.9pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1058" type="#_x0000_t75" style='width:237.75pt;height:189pt'>
| |
| - | <v:imagedata src="Background.files/image040.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=317 height=252
| |
| - |
| |
| - | src="Background.files/image041.gif" v:shapes="_x0000_i1058"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.5pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:7.1pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span class=GramE><span style='font-size:10.0pt;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene</span></span><span
| |
| - |
| |
| - | style='font-size:10.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'> in cells (pellet from 20mL LB culture
| |
| - | at <st1:chmetcnv TCSC="0" NumberType="1" Negative="False" HasSpace="False"
| |
| - |
| |
| - | SourceValue="37" UnitName="ﾰC" w:st="on">37°C</st1:chmetcnv> for 24h)
| |
| - | was extracted with 500μL acetone. 100μL acetone extract was diluted
| |
| - | with 100μL water and loaded on to MicroPlate Reader for a full spectrum
| |
| - | scan. Value normalised with blank (50% acetone) and OD<sub>600</sub>. Pure
| |
| - | carotene (from Invitrogen) was added as standard. Each graph shows average of
| |
| - | three readings.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=259 valign=top style='width:194.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:263.9pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1059" type="#_x0000_t75"
| |
| - | style='width:183.75pt;height:149.25pt;mso-wrap-distance-left:0;
| |
| - | mso-wrap-distance-right:0' o:allowoverlap="f" filled="t">
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image042.emz" o:title="" cropbottom="1187f"
| |
| - | cropright="15343f"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=245 height=199
| |
| - |
| |
| - | src="Background.files/image043.gif" v:shapes="_x0000_i1059"><![endif]><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1060" type="#_x0000_t75" style='width:183.75pt;height:136.5pt;
| |
| - | mso-wrap-distance-left:0;mso-wrap-distance-right:0' wrapcoords="-62 0 -62 21502 21600 21502 21600 0 -62 0"
| |
| - | filled="t">
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image044.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=245 height=182
| |
| - |
| |
| - | src="Background.files/image045.gif" v:shapes="_x0000_i1060"><![endif]></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p style='text-align:justify;text-justify:inter-ideograph;line-height:150%'><span
| |
| - |
| |
| - | style='font-size:11.0pt;line-height:150%;font-family:Arial'>To investigate the
| |
| - | induction of pigment production, we tested our device K<st1:chmetcnv TCSC="0"
| |
| - |
| |
| - | NumberType="1" Negative="False" HasSpace="True" SourceValue="274220"
| |
| - |
| |
| - | UnitName="in" w:st="on">274220 in</st1:chmetcnv> <i style='mso-bidi-font-style:
| |
| - |
| |
| - | normal'>E.coli</i> MG1655 and <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv>
| |
| - | arabinose was added to LB culture. Cell culture that was induced with arabinose
| |
| - | showed greater </span><span style='font-size:11.0pt;line-height:150%;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene
| |
| - | production than cell culture without arabinose, indicating the feasibility of
| |
| - | inducing pigment production. Using known amount of pure β-carotene to
| |
| - | construct a standard curve, we were able to estimate the amount of β-carotene
| |
| - | produced per unit volume of cell culture by our device.<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:303.15pt'>
| |
| - | <td width=307 valign=top style='width:230.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:303.15pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><span style='font-size:10.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1061" type="#_x0000_t75" style='width:3in;height:169.5pt'>
| |
| - | <v:imagedata src="Background.files/image046.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=288 height=226
| |
| - |
| |
| - | src="Background.files/image047.gif" v:shapes="_x0000_i1061"><![endif]></span><span
| |
| - |
| |
| - | style='font-size:10.5pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'><o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-right:6.7pt'><span style='font-size:10.0pt;
| |
| - |
| |
| - | font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene
| |
| - | in cells (pellet from 20mL LB culture at <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="37" UnitName="ﾰC"
| |
| - |
| |
| - | w:st="on">37°C</st1:chmetcnv> for 24h; <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv>
| |
| - | arabinose was used for induction) was extracted with 500μL acetone.
| |
| - | 100μL acetone extract was diluted with 100μL water and loaded on to
| |
| - | MicroPlate Reader for a full spectrum scan. Value normalised with blank (50%
| |
| - | acetone) and OD<sub>600</sub>. Pure carotene (Invitrogen) was added as
| |
| - | standard. Each graph shows average of three readings.<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=283 valign=top style='width:212.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:303.15pt'>
| |
| - | <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 align=left
| |
| - |
| |
| - | style='border-collapse:collapse;mso-table-layout-alt:fixed;mso-table-overlap:
| |
| - |
| |
| - | never;mso-table-lspace:9.0pt;margin-left:6.75pt;mso-table-rspace:9.0pt;
| |
| - |
| |
| - | margin-right:6.75pt;mso-table-anchor-vertical:paragraph;mso-table-anchor-horizontal:
| |
| - |
| |
| - | margin;mso-table-left:left;mso-table-top:162.45pt;mso-padding-alt:2.75pt 2.75pt 2.75pt 2.75pt'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;height:24.0pt'>
| |
| - | <td width=72 valign=top style='width:53.95pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-right:none;mso-border-top-alt:solid black .1pt;mso-border-left-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-bottom-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Samples<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=48 valign=top style='width:36.0pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-right:none;mso-border-top-alt:solid black .1pt;mso-border-left-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-bottom-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Abs. (unit)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=72 valign=top style='width:54.0pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-right:none;mso-border-top-alt:solid black .1pt;mso-border-left-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-bottom-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Carotene (μg /100μL
| |
| - | Acetone extract)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | <td width=85 valign=top style='width:63.5pt;border:solid black 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:24.0pt'>
| |
| - | <p class=TabellenInhalt style='mso-hyphenate:auto'><b><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>Total mass of carotene (μg
| |
| - | /mL-culture)<o:p></o:p></span></b></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:1;height:33.7pt'>
| |
| - | <td width=72 valign=top style='width:53.95pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:33.7pt'>
| |
| - | <p class=TabellenInhalt><span lang=DE style='font-size:9.0pt;font-family:
| |
| - |
| |
| - | Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman";
| |
| - |
| |
| - | mso-ansi-language:DE'>K<st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="True" SourceValue="274210" UnitName="in" w:st="on">274210
| |
| - | in</st1:chmetcnv> <i>E.coli</i> MG1655<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=48 valign=top style='width:36.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:33.7pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>0.4<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=72 valign=top style='width:54.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:33.7pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>6<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=85 valign=top style='width:63.5pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-right-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:33.7pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>6*5/20=1.5<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | <tr style='mso-yfti-irow:2;mso-yfti-lastrow:yes;height:45.45pt'>
| |
| - | <td width=72 valign=top style='width:53.95pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:45.45pt'>
| |
| - | <p class=TabellenInhalt><span style='font-size:9.0pt;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>K274220,
| |
| - | with <st1:chmetcnv TCSC="0" NumberType="1" Negative="False" HasSpace="False"
| |
| - |
| |
| - | SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv> ara. in <i>E.coli</i>
| |
| - | MG1655<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=48 valign=top style='width:36.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:45.45pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>0.3<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=72 valign=top style='width:54.0pt;border-top:none;border-left:
| |
| - |
| |
| - | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none;
| |
| - |
| |
| - | mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:solid black .1pt;
| |
| - |
| |
| - | padding:2.75pt 2.75pt 2.75pt 2.75pt;height:45.45pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>5<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=85 valign=top style='width:63.5pt;border:solid black 1.0pt;
| |
| - |
| |
| - | border-top:none;mso-border-left-alt:solid black .1pt;mso-border-bottom-alt:
| |
| - |
| |
| - | solid black .1pt;mso-border-right-alt:solid black .1pt;padding:2.75pt 2.75pt 2.75pt 2.75pt;
| |
| - |
| |
| - | height:45.45pt'>
| |
| - | <p class=TabellenInhalt align=right style='text-align:right'><span
| |
| - |
| |
| - | style='font-size:9.0pt;font-family:Arial;mso-fareast-font-family:Arial;
| |
| - |
| |
| - | mso-bidi-font-family:"Times New Roman"'>5*5/20=1.25<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:6.7pt;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm'><!--[if gte vml 1]><v:shape id="_x0000_i1062" type="#_x0000_t75"
| |
| - | style='width:219.75pt;height:149.25pt;mso-wrap-distance-left:0;
| |
| - | mso-wrap-distance-right:0'>
| |
| - | <v:fill color2="black"/>
| |
| - | <v:imagedata src="Background.files/image048.emz" o:title=""/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=293 height=199
| |
| - |
| |
| - | src="Background.files/image049.gif" v:shapes="_x0000_i1062"><![endif]></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <h3><u>Discussion of results<o:p></o:p></u></h3>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>Despite
| |
| - | relatively poor performance in <i style='mso-bidi-font-style:normal'>E.coli</i>
| |
| - | TOP10, our pigment-producing devices gave encouraging results in <i
| |
| - |
| |
| - | style='mso-bidi-font-style:normal'>E.coli</i> MG1655. Simply by overnight
| |
| - | incubation, the colonies were able to show visible colour changes. Since the
| |
| - | precursors were already present in normal <i style='mso-bidi-font-style:normal'>E.coli</i>,
| |
| - | no additional growth medium or special treatment was needed. This paves the way
| |
| - | to developing a signalling system that is quick to produce visual outputs.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial'>The
| |
| - | acetone extraction of carotenoid provides a low-cost method for quantitative
| |
| - | measurement. In the above experiments, the results of photo-spectrometry were
| |
| - | consistent with characteristic absorbance wavelengths of lycopene (around
| |
| - | 475nm) and </span><span style='font-size:11.0pt;line-height:150%;font-family:
| |
| - |
| |
| - | Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>β-carotene
| |
| - | (around 450nm), as noted in literature. In the case of β-carotene, the
| |
| - | absorbance profile of our β-carotene-producing device (K274210) matched
| |
| - | that of pure carotene standards, confirming that our device was functional in
| |
| - | the cells.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>When we
| |
| - | overlay the graphs of our two devices (constitutive), we obtained the following
| |
| - | result:<o:p></o:p></span></p>
| |
| - |
| |
| - | <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
| |
| - |
| |
| - | style='border-collapse:collapse;border:none;mso-border-alt:solid windowtext .5pt;
| |
| - |
| |
| - | mso-yfti-tbllook:480;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh:
| |
| - |
| |
| - | .5pt solid windowtext;mso-border-insidev:.5pt solid windowtext'>
| |
| - | <tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes;mso-yfti-lastrow:yes;
| |
| - |
| |
| - | height:186.4pt'>
| |
| - | <td width=379 valign=top style='width:284.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:186.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;text-align:justify;text-justify:inter-ideograph;
| |
| - |
| |
| - | line-height:150%'><span style='font-size:11.0pt;line-height:150%;font-family:
| |
| - |
| |
| - | Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><!--[if gte vml 1]><v:shape
| |
| - | id="_x0000_i1063" type="#_x0000_t75" style='width:273.75pt;height:189.75pt'>
| |
| - | <v:imagedata src="Background.files/image050.gif" o:title="234"/>
| |
| - | </v:shape><![endif]--><![if !vml]><img border=0 width=365 height=253
| |
| - |
| |
| - | src="Background.files/image050.gif" v:shapes="_x0000_i1063"><![endif]><o:p></o:p></span></p>
| |
| - | </td>
| |
| - | <td width=211 valign=top style='width:158.4pt;border:solid windowtext 1.0pt;
| |
| - |
| |
| - | border-left:none;mso-border-left-alt:solid windowtext .5pt;mso-border-alt:
| |
| - |
| |
| - | solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:186.4pt'>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>red: K274110/lycopene;<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>orange: K274210/β-carotene;<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>black: pure β-carotene;<o:p></o:p></span></p>
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:
| |
| - |
| |
| - | 6.0pt;margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;
| |
| - |
| |
| - | line-height:150%;font-family:Arial;mso-fareast-font-family:Arial;mso-bidi-font-family:
| |
| - |
| |
| - | "Times New Roman"'>blue: untransformed <i style='mso-bidi-font-style:normal'>E.coli</i>
| |
| - | MG1655 (negative control)<o:p></o:p></span></p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>Lycopene
| |
| - | and β-carotene showed characteristic absorbance profiles, with different
| |
| - | maximum absorbance wavelengths. It was tempted to think that by converting
| |
| - | lycopene to β-carotene (e.g. by expressing CrtY gene), we would be able to
| |
| - | see a “shift” in the peaks of maximum absorbance. However, in the bar charts
| |
| - | above it was noted that lycopene and β-carotene had significant absorbance
| |
| - | at both 456nm and 474nm, albeit with different ratios (i.e. lycopene absorbed
| |
| - | more at 474nm; β-carotene absorbed more at 456nm). Therefore, measurement
| |
| - | at a single wavelength was insufficient to determine the identity of the
| |
| - | carotenoid; we might need to use “456nm:474nm” ratio instead. It would be
| |
| - | interesting to see the effect of mixing different amounts of lycopene and β-carotene,
| |
| - | and whether the “456nm:474nm” ratio can indicate the relative percentage of
| |
| - | lycopene and β-carotene. In the current project, we were unable to obtain
| |
| - | pure lycopene due to high cost of purchase.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>In terms
| |
| - | of inducing β-carotene production, <st1:chmetcnv TCSC="0" NumberType="1"
| |
| - |
| |
| - | Negative="False" HasSpace="False" SourceValue="1" UnitName="mm" w:st="on">1mM</st1:chmetcnv>
| |
| - | arabinose, acting on Pbad promoter, was able to produce a level of synthesis
| |
| - | close to that under constitutive promoter. In the non-induced population, there
| |
| - | was probably a small degree of “leakiness” and basal level of gene expression
| |
| - | (leading to some β-carotene synthesis). In future experiments, it might be
| |
| - | helpful to move the pigment-producing devices to low copy plasmids and reduce
| |
| - | the effect of such basal expression.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'>One
| |
| - | possible design using our devices is to construct a system where lycopene is
| |
| - | constitutively produced and CrtY, which converts red lycopene to orange β-carotene,
| |
| - | is controlled by external stimulus. Hence, the red cells will turn orange upon
| |
| - | sensing the specific external stimulus, and this change in colour serves as the
| |
| - | signal output that can be detected visually and measured quantitatively at low
| |
| - | cost.<o:p></o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;text-align:justify;text-justify:inter-ideograph;line-height:
| |
| - |
| |
| - | 150%'><span style='font-size:11.0pt;line-height:150%;font-family:Arial;
| |
| - |
| |
| - | mso-fareast-font-family:Arial;mso-bidi-font-family:"Times New Roman"'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'><o:p> </o:p></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - |
| |
| - | margin-left:0cm;line-height:150%'><span style='font-size:11.0pt;line-height:
| |
| - |
| |
| - | 150%;font-family:Arial'><o:p> </o:p></span></p>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | </body>
| |
| - |
| |
| - | </html>
| |
| | | | |
| | <!--Do not remove the first and last lines in this page!-->{{Template:CambridgeBottom}} | | <!--Do not remove the first and last lines in this page!-->{{Template:CambridgeBottom}} |
"
(rbs + CrtE)
