Team:Cambridge/Project/CA02

From 2009.igem.org

(Difference between revisions)
(Create our own devices by standard assembly)
(Create our own devices by standard assembly)
Line 73: Line 73:
* Put the above two constructs under constitutive promoter (R0011) and arabinose-induced promoter/Pbad (I0500) and test the effect on colour production.
* Put the above two constructs under constitutive promoter (R0011) and arabinose-induced promoter/Pbad (I0500) and test the effect on colour production.
-
<table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0
+
{| style="color:#CCC; background-color:#3D5089;" cellpadding="5" cellspacing="0" border="1"
 +
!
 +
! Lycopene-producing device
 +
! β-carotene-producing device
 +
|- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1"
 +
|Constituent Biobricks (already in the registry)
 +
|Edinburgh 08
 +
|[[Image:Icon translational unit.png]] (rbs + CrtE)
 +
|insert: 922bp, backbone: 2079bp, total: 3001bp.
 +
|- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1"
 +
|K118006
 +
|Edinburgh 08
 +
|[[Image:Icon translational unit.png]] (rbs + CrtB)
 +
|insert: 949bp, backbone: 2079bp, total: 3028bp.
 +
|- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1"
 +
|K118005
 +
|Edinburgh 08
 +
|[[Image:Icon translational unit.png]] (rbs + CrtI)
 +
|insert: 1498bp, backbone: 2079bp, total: 3577bp.
 +
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)
 +
|insert: 1162bp, backbone: 2079bp, total: 3241bp.
 +
|- 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)
 +
|insert: 5412bp, backbone: 2079bp, total: 7491bp.
 +
2 PstI sites in CrtI removed.
 +
|- style="color:#333; background-color:#FFFFFF;" cellpadding="6" cellspacing="0" border="1"
 +
|}
-
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>&nbsp;</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'>&#946;-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>&nbsp;</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>
 
<!--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}}

Revision as of 01:59, 18 October 2009


Carotenoids

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:

Registry Code Team Sequence Description Size of insert
K118014 Edinburgh 08 Icon translational unit.png (rbs + CrtE) 922bp
K118006 Edinburgh 08 Icon translational unit.png (rbs + CrtB) 949bp
K118005 Edinburgh 08 Icon translational unit.png (rbs + CrtI) 1498bp. 2 PstI sites removed.
K118013 Edinburgh 08 Icon translational unit.png (rbs + CrtY) 1162bp
K152005 Guelph 08 Icon translational unit.pngIcon translational unit.pngIcon translational unit.pngIcon translational unit.pngIcon reporter.png

(rbs+CrtE) + (rbs+CrtB) + (rbs+CrtI) + (rbs+CrtY) + (rbs+GFP)

5412bp.2 PstI sites in CrtI removed.


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).


During our preliminary research, we encountered a few issues:

  • 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.
  • 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.
  • 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.


Create our own devices by standard assembly

Given the information at hand, we decided to adopt the following general strategies:

  • Create a lycopene-producing device (with enzymes CrtE, CrtB and CrtI) by standard assembly of Biobricks K118014, K118006 and K118005.
  • Create a β-carotene-producing device (with enzymes CrtE, CrtB, CrtI and CrtY) by standard assembly of Biobricks K118014, K118006, K118005 and K118013.
  • Put the above two constructs under constitutive promoter (R0011) and arabinose-induced promoter/Pbad (I0500) and test the effect on colour production.
Lycopene-producing device β-carotene-producing device
Constituent Biobricks (already in the registry) Edinburgh 08 Icon translational unit.png (rbs + CrtE) insert: 922bp, backbone: 2079bp, total: 3001bp.
K118006 Edinburgh 08 Icon translational unit.png (rbs + CrtB) insert: 949bp, backbone: 2079bp, total: 3028bp.
K118005 Edinburgh 08 Icon translational unit.png (rbs + CrtI) insert: 1498bp, backbone: 2079bp, total: 3577bp.

2 PstI sites removed.

K118013 Edinburgh 08 Icon translational unit.png (rbs + CrtY) insert: 1162bp, backbone: 2079bp, total: 3241bp.
K152005 Guelph 08 Icon translational unit.pngIcon translational unit.pngIcon translational unit.pngIcon translational unit.pngIcon reporter.png

(rbs+CrtE) + (rbs+CrtB) + (rbs+CrtI) + (rbs+CrtY) + (rbs+GFP)

insert: 5412bp, backbone: 2079bp, total: 7491bp.

2 PstI sites in CrtI removed.


Cambridge Sponsor Logo1.pngCambridge Sponsor Logo2.pngCambridge Sponsor Logo3.pngCambridge Sponsor Logo4.pngCambridge Sponsor Logo5.pngCambridge Sponsor Logo8.pngCambridge Sponsor Logo6.pngCambridge Sponsor Logo7.pngCambridge Sponsor Logo9.pngCambridge Sponsor Logo10.pngCambridge Sponsor Logo11.pngCambridge Sponsor Logo12.pngCambridge Sponsor Logo14.pngCambridge Sponsor Logo13.pngCambridge Sponsor Logo15.pngCambridge Sponsor Logo16.pngCambridge Sponsor Logo17.pngCambridge Sponsor Logo18.pngCambridge Sponsor Logo19.pngBmglab.jpg