Team:UC Davis/Parts

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style="color: rgb(255, 255, 153);"><span
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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/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
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><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: &quot;Times New Roman&quot;,&quot;serif&quot;;"></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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
 +
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="color: rgb(255, 255, 153);"><span
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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/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="color: rgb(255, 255, 153);"><span
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style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
 +
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>&nbsp;<a
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><small>
 +
</small></big></big></big></big></span></b>
 +
<hr style="width: 100%; height: 2px;">
 +
<div style="text-align: left;"><big><big><b
 +
style="color: rgb(0, 0, 0); text-decoration: underline;">Parts:</b></big></big><b
 +
style="color: rgb(0, 0, 0);">&nbsp;&nbsp; <br>
 +
<br>
 +
Parts related to
 +
secretion:&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
Parts related to pH sensor:<br>
 +
</b>
 +
<table
 +
style="text-align: left; width: 100%; background-color: rgb(255, 255, 0);"
 +
border="2" cellpadding="2" cellspacing="2">
 +
<tbody>
 +
<tr>
 +
<td style="vertical-align: top;">Proteins:<br>
 +
</td>
 +
<td style="vertical-align: top;">Promoters:<br>
 +
</td>
 +
<td style="vertical-align: top;">Others:<br>
 +
</td>
 +
<td style="vertical-align: top;">Proteins:<br>
 +
</td>
 +
<td style="vertical-align: top;">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
Promoters:<br>
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top;">
 +
<ul>
 +
<li><a href="#INPNC">INPNC</a></li>
 +
<li><a href="#OmpA">OmpA</a></li>
 +
<li><a href="#Luciferase">Luciferase</a></li>
 +
<li><a href="#GFP">GFP</a></li>
 +
</ul>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<ul>
 +
<li><a href="#LacI">LacI</a></li>
 +
</ul>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<ul>
 +
<li><a href="#SS">SS:signal sequence</a></li>
 +
<li><a href="#His">6-His-Tag</a></li>
 +
<li><a href="#RBS">RBS</a></li>
 +
<li><a href="#Terminator">Terminator</a></li>
 +
</ul>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<ul>
 +
<li>ChvI</li>
 +
<li>ChvG</li>
 +
</ul>
 +
</td>
 +
<td style="vertical-align: top;">
 +
<ul>
 +
<li><a href="#ChvI_promoter">ChvI promoter</a></li>
 +
<li><a href="#katA">KatA promoter</a></li>
 +
<li><a href="#aopB">AopB promoter</a></li>
 +
<li><a href="#PhoA">PhoA
 +
promoter</a>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
&nbsp;&nbsp; </li>
 +
<li>I<a href="#impA">mpA promoter</a></li>
 +
</ul>
 +
</td>
 +
</tr>
 +
</tbody>
 +
</table>
 +
<b style="color: rgb(0, 0, 0);">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
</b><br>
 +
<hr style="width: 100%; height: 2px;">
 +
<p class="MsoNormal" style=""><b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><a
 +
name="INPNC"></a>INPNC</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<p class="MsoNormal" style=""><i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">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: &quot;Times New Roman&quot;,&quot;serif&quot;;">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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;">OmpA</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
 +
</span><i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
 +
have modified this protein to Biobrick standard, Tom Knights
 +
Standard. </span></i><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><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: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:</span></i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
“<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>RBS</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
 +
</span><i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
 +
more information go to:</span></i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>Terminator</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
 +
</span><i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
 +
more information go to:</span></i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>GFP</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>Luciferase</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;">LacI</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;">SS</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
This signal sequence, when placed between INPNC, contains a
 +
cleavable site that allows the target fusion protein to ‘secrete’ from
 +
INPNC.&nbsp; We will do the same with OmpA.&nbsp; <u1:p></u1:p></span><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<p class="MsoNormal" style=""><i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
 +
have modified this protein to Biobrick standard, Tom Knights
 +
Standard.</span></i><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;">6-His
 +
Tag</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
 +
</span><i><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;">ChvI
 +
promoter</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<hr align="center" size="2" width="100%"></span></div>
 +
<p class="MsoNormal" style=""><b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><a
 +
name="katA"></a>KatA promoter</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
: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&nbsp; 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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;">AopB
 +
promoter</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>PhoA
 +
promoter</span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
 +
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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
 +
<div class="MsoNormal" style="text-align: center;" align="center"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
 +
<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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>ImpA
 +
promoter: </span></b><span
 +
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">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: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></u></b></p>
 +
<p class="MsoNormal"><span
 +
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p>&nbsp;</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: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></small></div>
</div>
</div>
 +
</body>
</html>
</html>
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===Note===
 
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If you choose to include a '''Parts Submitted to the Registry''' page, please list your parts here.  This is not necessary but it may be a nice list to keep track of.
 

Revision as of 21:40, 25 September 2009

kjaBD K

 
Parts:  

Parts related to secretion:                                                                                                                            Parts related to pH sensor:
Proteins:
Promoters:
Others:
Proteins:
      Promoters:
  • ChvI
  • ChvG
                   

INPNC: Ice-nucleation protein (INP) from Pseudomonas Syringae was suggested to be used for display of foreign proteins on the surface of E.coli(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 E.coli(9).

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.In our project we are intending to harness and make use of this feature by fusing a specific protein to it. We have modified this protein to Biobrick standard, Tom Knights Standard.


OmpA: OmpA is one of the proteins on the outer membrane of E.coli (13). OmpA has been found to be useful as utilizable fusion part that can fuse our protein to and display on the surface of E.coli. 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.
We have modified this protein to Biobrick standard, Tom Knights Standard.

Note:It has remained essentially unknown how proteins of Escherichia coli outer membrane are sorted and incorporated into this membrane” (10)
For more information go to:
http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836


RBS: Ribosome Binding site number 32 (BBa_J61132) from the registry is being used in our secretion system.
For more information go to: http://partsregistry.org/wiki/index.php/Part:BBa_J61132


Terminator: We are using BBa_B0015, a double terminator, as our terminator in both our secretion and pH system.
For more information go to: http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015


GFP: We are using Green Fluorescent Protein as a reporter that also serves as a small protein in testing our secretion system.


Luciferase: Luciferase is a firefly protein that also fluoresces, so it serves as a reporter as well as a testable large protein.
More can be found in:


LacI: One inducible Promoter which was found in the part registry.
More can be found in:
http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010


SS: 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. 

We have modified this protein to Biobrick standard, Tom Knights Standard.


6-His Tag: The 6-Histidine Tag serves as a tag for Western Blotting if our fluorescent reporters are not expressed as highly as we would like.
Note: We are using this tag, just in case if the GFP or Luciferase does not work under a plate reader.


ChvI promoter: 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.


KatA promoter :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 Agrobacterium tumorigenesis (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.


AopB promoter: 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 Agrobacterium tumorigenesis (2). Therefore, we have chosen this gene to be one of our candidates to complete our pH sensor device.


PhoA promoter: 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.


ImpA promoter: 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.