@@ Line 1: / Line 1: @@
-<html>
-<head>
-<meta content="text/html;charset=ISO-8859-1" http-equiv="Content-Type">
-<title>Model 1</title>
-</head>
-<body>
-<div style="text-align: center;"><span
-style="color: rgb(255, 255, 153);"><span
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><span
-style="font-weight: bold;">=</span></span></span><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-href="https://2009.igem.org/Team:UC_Davis"><img alt=""
-src="https://static.igem.org/mediawiki/2009/a/a4/UCDAVIS_PIC3.png"
-style="border: 0px solid ; width: 83px; height: 36px;"></a> </big></big></big></big></span></b><a
-href="https://2009.igem.org/Team:UC_Davis/About_Us"><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><img
-alt="" src="https://static.igem.org/mediawiki/2009/d/d1/UCDAVIS_PIC4.png"
-style="border: 0px solid ; width: 82px; height: 36px;"></big></big></big></big></span></b></a><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big>
-</big></big></big></big></span></b><b style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-href="https://2009.igem.org/Team:UC_Davis/Project"><img alt=""
-src="https://static.igem.org/mediawiki/2009/b/b9/UCDAVIS_PIC8.png"
-style="border: 0px solid ; width: 78px; height: 36px;"></a> </big></big></big></big></span></b><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-href="https://2009.igem.org/Team:UC_Davis/Notebook"><img alt=""
-src="https://static.igem.org/mediawiki/2009/2/2f/UCDAVIS_PIC5.png"
-style="border: 0px solid ; width: 81px; height: 36px;"></a> </big></big></big></big></span></b><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b
-style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-href="https://2009.igem.org/Team:UC_Davis/Parts"><img alt=""
-src="https://static.igem.org/mediawiki/2009/a/a6/UCDAVIS_PIC6.png"
-style="border: 0px solid ; width: 78px; height: 37px;"></a></big></big></big></big></span></b><b
-style="color: rgb(0, 0, 0);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b></div>
-<hr style="width: 100%; height: 2px;"><big><big><big><span
-style="text-decoration: underline; font-weight: bold;"><small>Secretion
-Model 2:<br>
-<br>
-</small></span></big></big></big><big><span style="font-style: italic;">Click
-on an
-individual part for more information.<br>
-</span></big>
-<div style="text-align: center;"><big><big><big><span
-style="font-weight: bold;"><small><span
-style="text-decoration: underline; background-color: rgb(255, 255, 0);"><a
-href="#LacI"><img alt=""
-src="https://static.igem.org/mediawiki/2009/4/4e/UCDAVIS_PROMOTER_1.png"
-style="border: 0px solid ; width: 242px; height: 58px; background-color: rgb(255, 255, 255);"></a><a
-href="#RBS"><img alt=""
-src="https://static.igem.org/mediawiki/2009/1/15/UCDAVIS_RBS1.png"
-style="border: 0px solid ; width: 89px; height: 81px; background-color: rgb(255, 255, 255);"></a><a
-href="#INPNC"><img alt=""
-src="https://static.igem.org/mediawiki/2009/a/ae/UCDAVIS_INPNC.png"
-style="border: 0px solid ; width: 106px; height: 59px; background-color: rgb(255, 255, 255);"></a><a
-href="#SS"><img alt=""
-src="https://static.igem.org/mediawiki/2009/9/9d/UCDAVIS_SS1.png"
-style="border: 0px solid ; width: 94px; height: 69px; background-color: rgb(255, 255, 255);"></a><img
-style="width: 106px; height: 59px; background-color: rgb(255, 255, 255);"
-alt="" src="https://static.igem.org/mediawiki/2009/0/0c/UCDAVIS_GENE1.png"><a
-href="#His"><img alt=""
-src="https://static.igem.org/mediawiki/2009/c/c0/UCDAVIS_6HIS.png"
-style="border: 0px solid ; width: 134px; height: 66px; background-color: rgb(255, 255, 255);"></a><a
-href="#Terminator"><img alt=""
-src="https://static.igem.org/mediawiki/2009/c/ca/UCDAVIS_STOP.png"
-style="border: 0px solid ; width: 94px; height: 83px; background-color: rgb(255, 255, 255);"></a><br>
-</span></small></span></big></big></big><big><big><big><span
-style="font-weight: bold;"><small><span
-style="text-decoration: underline; background-color: rgb(255, 255, 0);"><br>
-</span>a.<a href="#LacI"><span style="text-decoration: underline;"><img
-alt=""
-src="https://static.igem.org/mediawiki/2009/4/4e/UCDAVIS_PROMOTER_1.png"
-style="border: 0px solid ; width: 200px; height: 52px;"></span></a><span
-style="text-decoration: underline; background-color: rgb(255, 255, 0);"><a
-href="#RBS"><img alt=""
-src="https://static.igem.org/mediawiki/2009/1/15/UCDAVIS_RBS1.png"
-style="border: 0px solid ; width: 69px; height: 63px; background-color: rgb(255, 255, 255);"></a><a
-href="#INPNC"><img alt=""
-src="https://static.igem.org/mediawiki/2009/a/ae/UCDAVIS_INPNC.png"
-style="border: 0px solid ; width: 106px; height: 51px; background-color: rgb(255, 255, 255);"></a><a
-href="#SS"><img alt=""
-src="https://static.igem.org/mediawiki/2009/9/9d/UCDAVIS_SS1.png"
-style="border: 0px solid ; width: 73px; height: 65px; background-color: rgb(255, 255, 255);"></a><a
-href="#GFP"><img alt=""
-src="https://static.igem.org/mediawiki/2009/7/7d/UCDAVIS_GFP.png"
-style="border: 0px solid ; width: 83px; height: 55px; background-color: rgb(255, 255, 255);"></a><a
-href="#His"><img alt=""
-src="https://static.igem.org/mediawiki/2009/c/c0/UCDAVIS_6HIS.png"
-style="border: 0px solid ; width: 110px; height: 58px; background-color: rgb(255, 255, 255);"></a><a
-href="#Terminator"><img alt=""
-src="https://static.igem.org/mediawiki/2009/c/ca/UCDAVIS_STOP.png"
-style="border: 0px solid ; width: 78px; height: 71px; background-color: rgb(255, 255, 255);"></a></span></small></span></big></big></big><br>
-<div style="text-align: center;"><big><big><big><span
-style="font-weight: bold;"><small>b.<span
-style="text-decoration: underline;"><a href="#LacI"><img alt=""
-src="https://static.igem.org/mediawiki/2009/4/4e/UCDAVIS_PROMOTER_1.png"
-style="border: 0px solid ; width: 195px; height: 52px;"></a><a
-href="#RBS"><img alt=""
-src="https://static.igem.org/mediawiki/2009/1/15/UCDAVIS_RBS1.png"
-style="border: 0px solid ; width: 68px; height: 63px;"></a><a
-href="#INPNC"><img alt=""
-src="https://static.igem.org/mediawiki/2009/a/ae/UCDAVIS_INPNC.png"
-style="border: 0px solid ; width: 105px; height: 51px;"></a><a
-href="#SS"><img alt=""
-src="https://static.igem.org/mediawiki/2009/9/9d/UCDAVIS_SS1.png"
-style="border: 0px solid ; width: 73px; height: 65px;"></a><a
-href="#Luciferase"><img alt=""
-src="https://static.igem.org/mediawiki/2009/9/98/UCDAVIS_Luciferase.png"
-style="border: 0px solid ; width: 87px; height: 49px;"></a><a
-href="#His"><img alt=""
-src="https://static.igem.org/mediawiki/2009/c/c0/UCDAVIS_6HIS.png"
-style="border: 0px solid ; width: 110px; height: 58px;"></a><a
-href="#Terminator"><img alt=""
-src="https://static.igem.org/mediawiki/2009/c/ca/UCDAVIS_STOP.png"
-style="border: 0px solid ; width: 78px; height: 71px;"></a></span></small></span></big></big></big><big><big><big><span
-style="font-weight: bold;"><small><span
-style="text-decoration: underline; background-color: rgb(255, 255, 0);"></span></small></span></big></big></big></div>
-</div>
-<div style="text-align: center;"><big><span style="font-style: italic;"></span></big><br>
-</div>
-<big><big><big><span
-style="text-decoration: underline; font-weight: bold;"></span></big></big></big>
-<hr style="width: 100%; height: 2px;">
-<p class="MsoNormal" style=""><b><span style=""><a name="INPNC"></a>INPNC:
-</span></b><span style="">The
-ice-nucleation protein (INP) from <i>Pseudomonas syringae</i> is used
-by its
-natural host to nucleate ice formation and is implicated in<i> P.
-syringae</i>
-associated pathogenesis<i>.&nbsp; </i>INP and a truncated derivative
-lacking
-the central domain (INPNC) have been used extensively for displaying
-proteins
-on the surface of <i>E. coli (7)</i>.&nbsp; For instance, AldO and
-PhaZ1 have
-been successfully displayed on the surface of <i>E. coli </i>using
-INPNC (7,
-). <br>
-<u1:p></u1:p>Park <i>et al.</i> have shown that INPNC when fused to
-the <i>phaZ1</i>
-gene, including its signal sequence, can serve as a suitable surface
-delivery
-and secretion device of the otherwise toxic <i>phaZ1</i> gene product<i>
-</i>(15).&nbsp;
-<u1:p></u1:p>This part was synthesized by Mr. Gene (Regensburg,
-Germany) with
-codon optimization and subsequently transferred into vector (<span
-style="background: rgb(255, 255, 51) none repeat scroll 0% 50%; color: black; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span
-style="-moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial; background-attachment: scroll;">part
-name for AK vector).&nbsp;</span></span> As it is expected that this
-part will be used in the context of the fusion protein, the prefix and
-suffix
-for this part are consistent with the <i>BBF RCF-12</i>
-standard.&nbsp; <br>
-<u1:p></u1:p>We have proposed to build and test a general protein
-secretion
-system modeled after that developed by Park <i>et al. </i>in which a
-fusion of
-INPNC and the signal sequence from the <i>phaZ1</i> gene are used to
-secrete
-any target protein.&nbsp; <br>
-<i><u1:p></u1:p>We have modified this protein to be consistent with BBF
-RFC-12
-Standard. We have submitted this part to the parts registry as part </i><a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_K265008"><i><span
-style="color: rgb(0, 0, 153);">BBa_K265008</span></i></a><i>.</i> <o:p></o:p></span></p>
-<u2:p></u2:p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal" style=""><b><span style=""><a name="OmpA"></a>OmpA</span></b><span
-style="">: OmpA
-is one of the proteins on the outer membrane of <i>E. coli</i> (13),it
-is used
-as a displaying fusion protein on the cell surface . This part has
-already been
-documented on the parts registry; however, it has not been tested as a
-compnent
-of secretion system (via fusion with a target protein linked with a
-cleavable
-signal sequence) <i><br>
-<u1:p></u1:p>We have modified this protein to be consistent with BBF
-RFC-12 Standard.<br>
-Note: “It has remained essentially unknown how proteins of E. coli
-outer
-membrane are sorted and incorporated into this membrane” (10)</i> <i><br>
-For more information go to:<a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_K103006">
-BBa_K103006</a></i>
-<o:p></o:p></span></p>
-<u2:p></u2:p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal" style=""><b><span style=""><a name="RBS"></a>RBS</span></b><span
-style="">:&nbsp; Ribosome
-Binding site number 32 (BBa_J61132) from the registry is being used in
-our
-secretion system. <u2:p></u2:p><br>
-<i>For more information go to:</i> <a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><i>BBa_J61132</i></a><o:p></o:p></span></p>
-<u2:p></u2:p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal" style=""><b><span style=""><a name="Terminator"></a>Terminator</span></b><span
-style="">: We are
-using BBa_B0015, a double terminator, as our terminator in both our
-secretion
-and pH system.<u2:p></u2:p><br>
-<i>For more information go to:</i> <a
-href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015"><i>BBa_B0015</i></a>
-<o:p></o:p></span></p>
-<u2:p></u2:p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal" style=""><b><span style=""><a name="GFP"></a>GFP</span></b><span
-style="">: We are
-using Green Fluorescent Protein as a reporter that also serves as a
-small
-protein in testing our secretion system.<br>
-<i>For more informaiton go to: </i><a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_K265003"><i>BBa_K265003</i></a><i>&nbsp;</i><u2:p></u2:p><o:p></o:p></span></p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal" style=""><b><span style=""><a name="Luciferase"></a>Luciferase</span></b><span
-style="">: Luciferase
-is a firefly protein that also fluoresces, so it serves as a reporter
-as well
-as a testable large protein.<br>
-<i>For more information go to: <a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_I712019">BBa_1712019</a></i>
-<u2:p></u2:p><o:p></o:p></span></p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal" style=""><b><span style=""><a name="LacI"></a>LacI</span></b><span
-style="">: One
-inducible Promoter which was found in the part registry.<br>
-<i>For more information go to:<a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_R0010">
-BBa_R0010</a><br style="">
-<!--[endif]--></i><o:p></o:p></span></p>
-<u2:p></u2:p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal" style=""><b><span style=""><a name="SS"></a>SS</span></b><span
-style="">:The
-signal sequence (SS) for the <i>phaZ1 </i>gene product of <i>Paucimonas
-lemoignei</i>, a polyhydroxybutyrate depolymerase (15).&nbsp; In the
-native
-protein the signal sequence is cleaved between residues Ala37 and
-Leu38.&nbsp;
-Park <i>et al. </i>have showed that the fusion of the complete <i>phaZ1
-</i>gene
-(including SS) and a truncated ice nucleation protein from <i>Pseudomonas
-syringae</i> (<a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_K265008"><i><span
-style="color: rgb(0, 0, 153);">BBa_K265008</span></i></a>), could lead
-to stable
-expression and secretion of the <i>phaZ1</i> gene product.&nbsp; <br>
-<u1:p></u1:p>We propose that the signal sequence might be generally
-useful as a
-cleavage tag in secretion systems that include a membrane anchor
-component,
-such as INPNC (<a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_K265008"><i><span
-style="color: rgb(0, 0, 153);">BBa_K265008</span></i></a>) or OmpA (<i><a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_K103006">BBa_K103006</a>).<span
-style="color: rgb(0, 41, 57);"> </span></i>The proposed constructs
-would consists of a
-membrane anchor (INPNC or OmpA) followed by the cleavable signal
-sequence and
-finally a target protein marked for secretion.&nbsp; <br>
-<i><u1:p></u1:p>Since we expect that this part will be used in the
-context of a
-fusion protein, we have modified this protein to be consistent with BBF
-RFC-12
-Standard. We have submitted this part to the part registry as part </i><a
-href="http://partsregistry.org/wiki/index.php/Part:BBa_K265002"><i><span
-style="color: rgb(0, 0, 153);">BBa_K265002</span></i></a>. <o:p></o:p></span></p>
-<u2:p></u2:p>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
-style="">
-<hr align="center" size="2" width="100%"></span></div>
-<p class="MsoNormal"><b><span style=""><a name="Tag"></a>6-His Tag</span></b><span
-style="">:The 6-Histidine Tag serves as a tag for
-Western Blotting if our fluorescent reporters are not expressed as
-highly as we
-would like. <u2:p></u2:p><br>
-<i>Note: We are using this tag, just in case if the GFP or Luciferase
-does not
-work under a plate reader.</i></span></p>
-<p class="MsoNormal"><span style=""></span></p>
-<hr style="width: 100%; height: 2px;">
-<p class="MsoNormal"><span style="">&nbsp;<o:p></o:p></span></p>
-<p class="MsoNormal"><br>
-<span
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span><b><u><span
-style="font-size: 12pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></u></b></p>
-</body>
-</html>

Team:UC Davis/Adding secretion/model 4

From 2009.igem.org

Latest revision as of 16:44, 21 October 2009