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- | <title>kjaBD K</title>
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- | <div style="text-align: center;"><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/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="font-size: 13.5pt; font-family: "Times New Roman","serif";"><big><big><big><big>
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- | </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=""
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- | 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="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/Notebook1"><img alt=""
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- | src="https://static.igem.org/mediawiki/2009/2/2f/UCDAVIS_PIC5.png"
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- | style="border: 0px solid ; width: 81px; height: 36px;"></a> </big></big></big></big></span></b><b
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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/Part1"><img alt=""
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- | 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> <a
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- | href="https://2009.igem.org/Team:UC_Davis/Contact1"><img alt=""
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- | src="https://static.igem.org/mediawiki/2009/1/1d/UCDAVIS_PIC7.png"
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- | style="border: 0px solid ; width: 83px; 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";"><big><big><big><big><small>
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- | </small></big></big></big></big></span></b>
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- | <hr style="width: 100%; height: 2px;">
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- | <div style="text-align: left;"><big><big><b
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- | style="color: rgb(0, 0, 0); text-decoration: underline;">Parts:</b></big></big><b
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- | style="color: rgb(0, 0, 0);"> <br>
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- | <br>
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- | Parts related to
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- | secretion:
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- | Parts related to pH sensor:<br>
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- | </b>
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- | <table
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- | style="text-align: left; width: 100%; background-color: rgb(255, 255, 0);"
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- | border="2" cellpadding="2" cellspacing="2">
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- | <tbody>
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- | <tr>
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- | <td style="vertical-align: top;">Proteins:<br>
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- | </td>
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- | <td style="vertical-align: top;">Promoters:<br>
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- | </td>
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- | <td style="vertical-align: top;">Others:<br>
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- | </td>
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- | <td style="vertical-align: top;">Proteins:<br>
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- | </td>
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- | <td style="vertical-align: top;">
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- | Promoters:<br>
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- | </td>
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- | </tr>
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- | <tr>
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- | <td style="vertical-align: top;">
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- | <ul>
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- | <li><a href="#INPNC">INPNC</a></li>
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- | <li><a href="#OmpA">OmpA</a></li>
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- | <li><a href="#Luciferase">Luciferase</a></li>
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- | <li><a href="#GFP">GFP</a></li>
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- | </ul>
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- | </td>
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- | <td style="vertical-align: top;">
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- | <ul>
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- | <li><a href="#LacI">LacI</a></li>
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- | </ul>
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- | </td>
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- | <td style="vertical-align: top;">
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- | <ul>
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- | <li><a href="#SS">SS:signal sequence</a></li>
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- | <li><a href="#His">6-His-Tag</a></li>
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- | <li><a href="#RBS">RBS</a></li>
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- | <li><a href="#Terminator">Terminator</a></li>
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- | </ul>
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- | </td>
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- | <td style="vertical-align: top;">
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- | <ul>
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- | <li>ChvI</li>
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- | <li>ChvG</li>
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- | </ul>
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- | </td>
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- | <td style="vertical-align: top;">
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- | <ul>
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- | <li><a href="#ChvI_promoter">ChvI promoter</a></li>
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- | <li><a href="#katA">KatA promoter</a></li>
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- | <li><a href="#aopB">AopB promoter</a></li>
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- | <li><a href="#PhoA">PhoA
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- | promoter</a>
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- | </li>
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- | <li>I<a href="#impA">mpA promoter</a></li>
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- | </ul>
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- | </td>
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- | </tr>
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- | </tbody>
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- | </table>
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- | <b style="color: rgb(0, 0, 0);">
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- | </b><br>
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- | <hr style="width: 100%; height: 2px;">
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- | <p class="MsoNormal" style=""><b><span
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- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><a
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- | name="INPNC"></a>INPNC</span></b><span
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- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
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- | Ice-nucleation protein (INP) from Pseudomonas Syringae was
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- | suggested to be used for display of foreign proteins on the surface of <i>E.coli</i>(7).Furthermore,
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- | researches have shown that an INP derivative constituting the N-and
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- | C-terminal
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- | domains can and has been used to display foreign proteins on the
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- | surface of <i>E.coli</i>(9).
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- | </span><span style="font-family: "Times New Roman","serif";"><o:p></o:p></span></p>
| + | |
- | <p class="MsoNormal" style=""><i><span
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- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">Note:
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- | A study has shown that "Ice- nucleation Protein (INP), an outer
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- | membrane
| + | |
- | protein from Pseudomonas syringae, is able to catalyze the ice crystal
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- | formation of supercooled water.</span></i><span
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- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">In
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- | our project we are intending to harness and make use of this feature by
| + | |
- | fusing
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- | 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
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- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">OmpA</span></b><span
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- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
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- | OmpA is one of the proteins on the outer membrane of <i>E.coli</i>
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- | (13). OmpA has been found to be useful as utilizable fusion part that
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- | can fuse
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- | our protein to and display on the surface of <i>E.coli</i>. This part
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- | has
| + | |
- | already been documented on the parts registry; however, it has not been
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- | tested
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- | 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><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="ChvI_promoter"></a><b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">ChvI
| + | |
- | promoter</span></b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
- | Gene fusion studies confirmed that ChvI gene was induced by
| + | |
- | acidic conditions (1). Also, it has been known to implicate in
| + | |
- | virulence (1).
| + | |
- | This gene is one of the candidates to be use in our biological pH
| + | |
- | sensor as a
| + | |
- | promoter.</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=""><b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><a
| + | |
- | name="katA"></a>KatA promoter</span></b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">
| + | |
- | :This
| + | |
- | Chromosomal gene is located on the linear chromosome (2) and it seems
| + | |
- | to be
| + | |
- | induced under an acidic environment as well as being involved in the <i>Agrobacterium
| + | |
- | tumorigenesis</i> (2).Research has suggested that ChvG is needed for
| + | |
- | "responsiveness of gene expression to low pH "(2). This gene
| + | |
- | has become a candidate to complete our pH sensor device from this
| + | |
- | evidence.</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="aopB"></a><b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">AopB
| + | |
- | promoter</span></b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
- | This Chromosomal gene located on the circular chromosome (2)
| + | |
- | encodes an outer member protein exposed on the bacterial cell surface
| + | |
- | (2).
| + | |
- | Also, ChvG was shown to be absolutely required for this gene expression
| + | |
- | (2)It
| + | |
- | seems to get induced under an acidic environment as well as being
| + | |
- | involved in
| + | |
- | the <i>Agrobacterium</i> <i>tumorigenesis </i>(2). Therefore, we
| + | |
- | have chosen
| + | |
- | this gene to be one of our candidates to complete our pH sensor device.</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="PhoA"></a><b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><u1:p></u1:p>PhoA
| + | |
- | promoter</span></b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">:
| + | |
- | There has been a suggestion that ChvI can activate AP activity by
| + | |
- | activating transcription of this gene, PhoA (3). Therefore, this gene
| + | |
- | has
| + | |
- | become one of our candidates to complete our pH sensor device.</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="impA"></a><b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";"><u1:p></u1:p>ImpA
| + | |
- | promoter: </span></b><span
| + | |
- | style="font-size: 13pt; line-height: 115%; font-family: "Times New Roman","serif";">Gene
| + | |
- | fusion studies confirmed that
| + | |
- | impA genes was induced by acidic conditions (1), therefore, this is one
| + | |
- | of our
| + | |
- | candidates to complete our pH sensor device.</span><b><u><span
| + | |
- | style="font-size: 12pt; line-height: 115%; font-family: "Times New Roman","serif";"><o:p></o:p></span></u></b></p>
| + | |
- | <p class="MsoNormal"><span
| + | |
- | style="font-family: "Times New Roman","serif";"><o:p> </o:p></span></p>
| + | |
- | <hr style="width: 100%; height: 2px;"><small style="font-weight: bold;"><span
| + | |
- | style="font-size: 18pt; line-height: 115%; font-family: "Times New Roman","serif";"></span></small></div>
| + | |
- | </div>
| + | |
- | </body>
| + | |
- | </html>
| + | |