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| - | <head>
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| - | <meta content="text/html;charset=ISO-8859-1" http-equiv="Content-Type">
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| - | <title>Model 1</title>
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| - | </head>
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| - | <body>
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| - | <div style="text-align: center;"><span
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-family: "Times New Roman","serif";"><span
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| - | style="font-weight: bold;">=</span></span></span><b
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"><big><big><big><big><a
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| - | href="https://2009.igem.org/Team:UC_Davis/homepage2"><img alt=""
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| - | src="https://static.igem.org/mediawiki/2009/a/a4/UCDAVIS_PIC3.png"
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| - | style="border: 0px solid ; width: 83px; height: 36px;"></a> </big></big></big></big></span></b><a
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| - | href="https://2009.igem.org/Team:UC_Davis/About_Us1"><b
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"><big><big><big><big><img
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| - | alt="" src="https://static.igem.org/mediawiki/2009/d/d1/UCDAVIS_PIC4.png"
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| - | style="border: 0px solid ; width: 82px; height: 36px;"></big></big></big></big></span></b></a><b
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"></span></b><b
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"></span></b><b
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"><big><big><big><big>
| + | |
| - | </big></big></big></big></span></b><b style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"><big><big><big><big><a
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| - | href="https://2009.igem.org/Team:UC_Davis/Project1"><img alt=""
| + | |
| - | src="https://static.igem.org/mediawiki/2009/b/b9/UCDAVIS_PIC8.png"
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| - | style="border: 0px solid ; width: 78px; height: 36px;"></a> </big></big></big></big></span></b><b
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"><big><big><big><big><a
| + | |
| - | href="https://2009.igem.org/Team:UC_Davis/Notebook1"><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
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"></span></b><b
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| - | style="color: rgb(255, 255, 153);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"><big><big><big><big><a
| + | |
| - | href="https://2009.igem.org/Team:UC_Davis/Part1"><img alt=""
| + | |
| - | src="https://static.igem.org/mediawiki/2009/a/a6/UCDAVIS_PIC6.png"
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| - | style="border: 0px solid ; width: 78px; height: 37px;"></a></big></big></big></big></span></b><b
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| - | style="color: rgb(0, 0, 0);"><span
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| - | style="font-size: 13.5pt; font-family: "Times New Roman","serif";"></span></b></div>
| + | |
| - | <hr style="width: 100%; height: 2px;"><big><big><big><span
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| - | style="text-decoration: underline; font-weight: bold;"><small>Secretion
| + | |
| - | Model
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| - | 1:<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="#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="#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="#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"><b><span
| + | |
| - | style="font-size: 18pt; line-height: 115%; font-family: "Times New Roman","serif";"><a
| + | |
| - | name="INPNC"></a></span></b><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">INPNC</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | Ice-nucleation protein (INP) from Pseudomonas Syringae was
| + | |
| - | suggested to be used for display of foreign proteins on the surface of <i>E.coli</i>(7).Furthermore,
| + | |
| - | researches have shown that an INP derivative constituting the N-and
| + | |
| - | C-terminal
| + | |
| - | domains can and has been used to display foreign proteins on the
| + | |
| - | surface of <i>E.coli</i>(9).
| + | |
| - | </span><span style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <p class="MsoNormal" style=""><i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">Note:
| + | |
| - | A study has shown that "Ice- nucleation Protein (INP), an outer
| + | |
| - | membrane
| + | |
| - | protein from Pseudomonas syringae, is able to catalyze the ice crystal
| + | |
| - | formation of supercooled water.</span></i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">In
| + | |
| - | our project we are intending to harness and make use of this feature by
| + | |
| - | fusing
| + | |
| - | a specific protein to it. <i>We have modified this protein to Biobrick
| + | |
| - | standard, Tom Knights Standard.</i></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal" style=""><a name="OmpA"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">OmpA</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | OmpA is one of the proteins on the outer membrane of <i>E.coli</i>
| + | |
| - | (13). OmpA has been found to be useful as utilizable fusion part that
| + | |
| - | can fuse
| + | |
| - | our protein to and display on the surface of <i>E.coli</i>. This part
| + | |
| - | has
| + | |
| - | already been documented on the parts registry; however, it has not been
| + | |
| - | tested
| + | |
| - | via fusion with a target protein linked with a cleavable signal
| + | |
| - | sequence.<u1:p></u1:p></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><br>
| + | |
| - | </span><i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">We
| + | |
| - | have modified this protein to Biobrick standard, Tom Knights
| + | |
| - | Standard. </span></i><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><u1:p></u1:p><o:p></o:p></span></p>
| + | |
| - | <p class="MsoNormal" style=""><i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">Note:</span></i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">
| + | |
| - | “<i>It has
| + | |
| - | remained essentially unknown how proteins of Escherichia coli outer
| + | |
| - | membrane
| + | |
| - | are sorted and incorporated into this membrane” (10)<br>
| + | |
| - | <u1:p></u1:p>For more information go to:</i> <a
| + | |
| - | href="http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836"><i><span
| + | |
| - | style="">http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836</span></i></a></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal" style=""><a name="RBS"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><u1:p></u1:p>RBS</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | Ribosome Binding site number 32 (BBa_J61132) from the registry is
| + | |
| - | being used in our secretion system.<u1:p></u1:p></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><br>
| + | |
| - | </span><i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">For
| + | |
| - | more information go to:</span></i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">
| + | |
| - | <a href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><i><span
| + | |
| - | style="">http://partsregistry.org/wiki/index.php/Part:BBa_J61132</span></i></a></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal" style=""><a name="Terminator"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><u1:p></u1:p>Terminator</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | We are using BBa_B0015, a double terminator, as our terminator in
| + | |
| - | both our secretion and pH system.<u1:p></u1:p></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><br>
| + | |
| - | </span><i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">For
| + | |
| - | more information go to:</span></i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">
| + | |
| - | <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015"><i><span
| + | |
| - | style="">http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015</span></i></a></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal" style=""><a name="GFP"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><u1:p></u1:p>GFP</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | We are using Green Fluorescent Protein as a reporter that also
| + | |
| - | serves as a small protein in testing our secretion system.</span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal" style=""><a name="Luciferase"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><u1:p></u1:p>Luciferase</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | Luciferase is a firefly protein that also fluoresces, so it
| + | |
| - | serves as a reporter as well as a testable large protein.<i><br>
| + | |
| - | More can be found in:</i></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal" style=""><a name="LacI"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">LacI</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | One inducible Promoter which was found in the part registry.<i><br>
| + | |
| - | More can be found in: </i><a
| + | |
| - | href="http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010"><i><span
| + | |
| - | style="">http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010</span></i></a></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal"><a name="SS"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">SS</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | This signal sequence, when placed between INPNC, contains a
| + | |
| - | cleavable site that allows the target fusion protein to ‘secrete’ from
| + | |
| - | INPNC. We will do the same with OmpA. <u1:p></u1:p></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <p class="MsoNormal" style=""><i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">We
| + | |
| - | have modified this protein to Biobrick standard, Tom Knights
| + | |
| - | Standard.</span></i><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
| - | <div class="MsoNormal" style="text-align: center;" align="center"><span
| + | |
| - | style="font-family: "Times New Roman","serif";">
| + | |
| - | <hr align="center" size="2" width="100%"></span></div>
| + | |
| - | <p class="MsoNormal" style=""><a name="His"></a><b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">6-His
| + | |
| - | Tag</span></b><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
| - | The 6-Histidine Tag serves as a tag for Western Blotting if our
| + | |
| - | fluorescent reporters are not expressed as highly as we would like.<u1:p></u1:p></span><span
| + | |
| - | style="font-family: "Times New Roman","serif";"><br>
| + | |
| - | </span><i><span
| + | |
| - | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">Note:
| + | |
| - | We are using this tag, just in case if the GFP or Luciferase
| + | |
| - | does not work under a plate reader.</span></i></p>
| + | |
| - | <b><i><span
| + | |
| - | style="font-size: 12pt; line-height: 115%; font-family: "Times New Roman","serif";"></span></i></b><big><big><big><span
| + | |
| - | style="text-decoration: underline; font-weight: bold;"></span></big></big></big>
| + | |
| - | </body>
| + | |
| - | </html>
| + | |
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