Team:UC Davis/Parts 1

From 2009.igem.org

(Difference between revisions)
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</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
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=""
href="https://2009.igem.org/Team:UC_Davis/homepage2"><img alt=""
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style="color: rgb(0, 0, 0);"><span
style="color: rgb(0, 0, 0);"><span
style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><small>
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><b
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</small></big></big></big></big></span></b>
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style="color: rgb(0, 0, 0);">&nbsp;&nbsp; </b>
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<hr style="width: 100%; height: 2px;">
<hr style="width: 100%; height: 2px;">
-
<div style="text-align: left;"><b
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<div style="text-align: left;"><big><big><b
-
style="color: rgb(0, 0, 0); text-decoration: underline;">Parts:</b><b
+
style="color: rgb(0, 0, 0); text-decoration: underline;">Parts:</b></big></big><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;&nbsp;&nbsp;&nbsp;
+
style="color: rgb(0, 0, 0);">&nbsp;&nbsp; <br>
<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>
</b>
-
<div style="text-align: left;"><b style="color: rgb(0, 0, 0);">Parts
+
<table
-
related to
+
style="text-align: left; width: 100%; background-color: rgb(255, 255, 0);"
-
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;
+
border="2" cellpadding="2" cellspacing="2">
-
Parts related to the biological pH sensor:</b><br>
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<tbody>
-
</div>
+
<tr>
-
<b style="color: rgb(0, 0, 0);">Protines:&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
<td style="vertical-align: top;">Proteins:<br>
-
&nbsp;
+
</td>
-
Promoters:&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
<td style="vertical-align: top;">Promoters:<br>
-
Others:&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;
+
</td>
-
Proteins:&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
<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>
Promoters:<br>
-
</b><a href="#INPNC">INPNC&nbsp;</a>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</td>
-
&nbsp;
+
</tr>
-
<a href="#LacI">LacI&nbsp;&nbsp;</a>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
<tr>
-
<a href="#SS">SS:signal
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<td style="vertical-align: top;">
-
sequence&nbsp;</a>&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;
+
<ul>
-
ChvI&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
<li><a href="#INPNC">INPNC</a></li>
-
<a href="#ChvI_promoter">ChvI promoter</a><br>
+
<li><a href="#OmpA">OmpA</a></li>
-
<a href="#OmpA">OmpA&nbsp;&nbsp;</a>&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;
+
<li><a href="#Luciferase">Luciferase</a></li>
-
<a href="#His">6-His-Tag</a>&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;
+
<li><a href="#GFP">GFP</a></li>
-
ChvG&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+
</ul>
-
<a href="#KatA">KatA promoter</a><br>
+
</td>
-
<a href="#Luciferase">Luciferase&nbsp;</a>&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;
+
<td style="vertical-align: top;">
-
<a href="#RBS">RBS</a>&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;
+
<ul>
-
<a href="#aopB">aopB promoter</a><br>
+
<li><a href="#LacI">LacI</a></li>
-
<a href="#GFP">GFP</a>&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;
+
</ul>
-
<a href="#Terminator">Terminator</a>&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;
+
</td>
-
<a href="#PhoA">phoA
+
<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;
promoter</a>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
-
&nbsp;&nbsp; <br>
+
&nbsp;&nbsp; </li>
-
&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;&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;
+
<li>I<a href="#impA">mpA promoter</a></li>
-
<a href="#impA">impA promoter</a><br>
+
</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;">
<hr style="width: 100%; height: 2px;">
-
<p class="MsoNormal"
+
<p class="MsoNormal" style=""><b><span
-
style="margin-bottom: 0.0001pt; line-height: normal;"><a name="INPNC"></a><span
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><a
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">INPNC:
+
name="INPNC"></a>INPNC</span></b><span
-
Ice-nucleation protein (INP) from Pseudomonas Syringae was suggested to
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
-
be used
+
Ice-nucleation protein (INP) from Pseudomonas Syringae was
-
for display of foreign proteins on the surface of <i>E.coli</i>(7).Furthermore,
+
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
researches have shown that an INP derivative constituting the N-and
C-terminal
C-terminal
domains can and has been used to display foreign proteins on the
domains can and has been used to display foreign proteins on the
surface of <i>E.coli</i>(9).
surface of <i>E.coli</i>(9).
-
<i><br>
+
</span><span style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
</i></span></p>
+
<p class="MsoNormal" style=""><i><span
-
<p class="MsoNormal"
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
-
style="margin-bottom: 0.0001pt; line-height: normal;"><span
+
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><i>Note:
+
A study has shown that "Ice- nucleation Protein (INP), an outer
A study has shown that "Ice- nucleation Protein (INP), an outer
-
membrane protein from Pseudomonas syringae, is able to catalyze the ice
+
membrane
-
crystal
+
protein from Pseudomonas syringae, is able to catalyze the ice crystal
-
formation of supercooled water.</i>In our project we are intending to
+
formation of supercooled water.</span></i><span
-
harness
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">In
-
and make use of this feature by fusing a specific protein to it. <i>We
+
our project we are intending to harness and make use of this feature by
-
have
+
fusing
-
modified this protein to Biobrick standard, Tom Knights Standard. </i><o:p></o:p></span></p>
+
a specific protein to it. <i>We have modified this protein to Biobrick
-
<p class="MsoNormal" style="line-height: normal;"><a name="OmpA"></a><span
+
standard, Tom Knights Standard.</i></span><span
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">OmpA:
+
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>
OmpA is one of the proteins on the outer membrane of <i>E.coli</i>
-
(13). OmpA
+
(13). OmpA has been found to be useful as utilizable fusion part that
-
has been found to be useful as utilizable fusion part that can fuse our
+
can fuse
-
protein
+
our protein to and display on the surface of <i>E.coli</i>. This part
-
to and display on the surface of <i>E.coli</i>. This part has already
+
has
-
been
+
already been documented on the parts registry; however, it has not been
-
documented on the parts registry; however, it has not been tested via
+
tested
-
fusion
+
via fusion with a target protein linked with a cleavable signal
-
with a target protein linked with a cleavable signal sequence.<o:p></o:p></span><br>
+
sequence.<u1:p></u1:p></span><span
-
<i><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
+
</span><i><span
-
have modified this protein to
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
-
Biobrick standard, Tom Knights Standard. </span></i><span
+
have modified this protein to Biobrick standard, Tom Knights
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+
Standard. </span></i><span
-
<p class="MsoNormal" style="line-height: normal;"><i><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p><o:p></o:p></span></p>
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:</span></i><span
+
<p class="MsoNormal" style=""><i><span
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> “<i>It
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:</span></i><span
-
has remained essentially unknown how proteins of
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
Escherichia coli outer membrane are sorted and incorporated into this
+
“<i>It has
-
membrane”
+
remained essentially unknown how proteins of Escherichia coli outer
-
(10)</i><o:p></o:p></span><i><span
+
membrane
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
+
are sorted and incorporated into this membrane” (10)<br>
-
For more information go to:</span></i><span
+
<u1:p></u1:p>For more information go to:</i> <a
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <a
+
href="http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836"><i><span
href="http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836"><i><span
-
style="color: blue;">http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836</span></i></a><br
+
style="">http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836</span></i></a></span><span
-
style="">
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
<!--[if !supportLineBreakNewLine]--><br style="">
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
<!--[endif]--><o:p></o:p></span></p>
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
<p class="MsoNormal" style="line-height: normal;"><a name="RBS"></a><span
+
<hr align="center" size="2" width="100%"></span></div>
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">RBS:
+
<p class="MsoNormal" style=""><a name="RBS"></a><b><span
-
Ribosome Binding site number 32 (BBa_J61132) from the registry is being
+
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
-
used in
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
-
our secretion system.<o:p></o:p></span><br>
+
Ribosome Binding site number 32 (BBa_J61132) from the registry is
-
<i><span
+
being used in our secretion system.<u1:p></u1:p></span><span
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
+
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
more information go to:</span></i><span
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <a
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><i><span
+
<a href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><i><span
-
style="color: blue;">http://partsregistry.org/wiki/index.php/Part:BBa_J61132</span></i></a><o:p></o:p></span></p>
+
style="">http://partsregistry.org/wiki/index.php/Part:BBa_J61132</span></i></a></span><span
-
<p class="MsoNormal" style="line-height: normal;"><a name="Terminator"></a><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Terminator:
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
We are using BBa_B0015, a double terminator, as our terminator in both
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
our
+
<hr align="center" size="2" width="100%"></span></div>
-
secretion and pH system.<o:p></o:p></span><br>
+
<p class="MsoNormal" style=""><a name="Terminator"></a><b><span
-
<i><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; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
-
more information go to:</span></i><a name="GFP:"></a><span
+
We are using BBa_B0015, a double terminator, as our terminator in
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <a
+
both our secretion and pH system.<u1:p></u1:p></span><span
-
href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015"><i><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
-
style="color: blue;">http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015</span></i></a><o:p></o:p></span></p>
+
</span><i><span
-
<p class="MsoNormal" style="line-height: normal;"><a name="GFP"></a><span
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">GFP:
+
more information go to:</span></i><span
-
We are using Green Fluorescent Protein as a reporter that also serves
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
as a
+
<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015"><i><span
-
small protein in testing our secretion system.<o:p></o:p></span></p>
+
style="">http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015</span></i></a></span><span
-
<p class="MsoNormal" style="line-height: normal;"><a name="Luciferase"></a><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Luciferase:
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
Luciferase is a firefly protein that also fluoresces, so it serves as a
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
reporter as well as a testable large protein.<i><br>
+
<hr align="center" size="2" width="100%"></span></div>
-
More can be found in<a name="LacI:"></a>:</i><o:p></o:p></span></p>
+
<p class="MsoNormal" style=""><a name="GFP"></a><b><span
-
<p class="MsoNormal" style="line-height: normal;"><a name="LacI"></a><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; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">LacI:
+
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>
One inducible Promoter which was found in the part registry.<i><br>
More can be found in: </i><a
More can be found in: </i><a
href="http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010"><i><span
href="http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010"><i><span
-
style="color: blue;">http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010</span></i></a><br>
+
style="">http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010</span></i></a></span><span
-
<br>
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
<a name="SS"></a>SS: This signal sequence, when placed between INPNC,
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
contains a
+
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
cleavable site that allows the target fusion protein to ‘secrete’ from
-
INPNC.&nbsp; We will do the same with OmpA.&nbsp; <o:p></o:p></span><br>
+
INPNC.&nbsp; We will do the same with OmpA.&nbsp; <u1:p></u1:p></span><span
-
<i><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
+
<p class="MsoNormal" style=""><i><span
-
have modified this protein to
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
-
Biobrick standard, Tom Knights Standard. </span></i><span
+
have modified this protein to Biobrick standard, Tom Knights
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+
Standard.</span></i><span
-
<p class="MsoNormal" style="line-height: normal;"><a name="His"></a><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">6-His
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
Tag: The 6-Histidine Tag serves as a tag for Western Blotting if our
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
fluorescent reporters are not expressed as highly as we would like.<o:p></o:p></span><br>
+
<hr align="center" size="2" width="100%"></span></div>
-
<i><span
+
<p class="MsoNormal" style=""><a name="His"></a><b><span
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">6-His
-
We are using this tag, just in
+
Tag</span></b><span
-
case if the GFP or Luciferase does not work under a plate reader.</span></i><span
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+
The 6-Histidine Tag serves as a tag for Western Blotting if our
-
<p class="MsoNormal" style="line-height: normal;"><span
+
fluorescent reporters are not expressed as highly as we would like.<u1:p></u1:p></span><span
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><a
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
-
name="ChvI_promoter"></a>ChvI promoter: Gene fusion studies
+
</span><i><span
-
confirmed that ChvI gene was induced by acidic conditions (1). Also, it
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
-
has
+
We are using this tag, just in case if the GFP or Luciferase
-
been known to implicate in virulence (1). This gene is one of the
+
does not work under a plate reader.</span></i><span
-
candidates to
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
be use in our biological pH sensor as a promoter.<br style="">
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
<!--[if !supportLineBreakNewLine]--><br style="">
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
<!--[endif]--><o:p></o:p></span></p>
+
<hr align="center" size="2" width="100%"></span></div>
-
<p class="MsoNormal" style="line-height: normal;"><a name="KatA"></a><span
+
<p class="MsoNormal" style=""><a name="ChvI_promoter"></a><b><span
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">KatA
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">ChvI
-
promoter :This Chromosomal gene is located on the linear chromosome (2)
+
promoter</span></b><span
-
and it
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
-
seems to be induced under an acidic environment as well as being
+
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
involved in
-
the <i>Agrobacterium tumorigenesis</i> (2).Research has suggested that
+
the <i>Agrobacterium</i> <i>tumorigenesis </i>(2). Therefore, we
-
ChvG is
+
have chosen
-
needed for "responsiveness of&nbsp; gene expression to low pH "(2).
+
this gene to be one of our candidates to complete our pH sensor device.</span><span
-
This gene has become a candidate to complete our pH sensor device from
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
this
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
evidence.<br style="">
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
<!--[if !supportLineBreakNewLine]--><br style="">
+
<hr align="center" size="2" width="100%"></span></div>
-
<!--[endif]--><o:p></o:p></span></p>
+
<p class="MsoNormal" style=""><a name="PhoA"></a><b><span
-
<p class="MsoNormal" style="line-height: normal;"><a name="aopB"></a><span
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>PhoA
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">aopB
+
promoter</span></b><span
-
promoter: This Chromosomal gene located on the circular chromosome (2)
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
-
encodes an
+
There has been a suggestion that ChvI can activate AP activity by
-
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.<o:p></o:p></span></p>
+
-
<p class="MsoNormal" style="line-height: normal;"><a name="PhoA"></a><span
+
-
style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">PhoA
+
-
promoter: There has been a suggestion that ChvI can activate AP
+
-
activity by
+
activating transcription of this gene, PhoA (3). Therefore, this gene
activating transcription of this gene, PhoA (3). Therefore, this gene
has
has
-
become one of our candidates to complete our pH sensor device.<o:p></o:p></span></p>
+
become one of our candidates to complete our pH sensor device.</span><span
-
<a name="impA"></a><span
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
-
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">impA
+
<div class="MsoNormal" style="text-align: center;" align="center"><span
-
promoter: Gene
+
style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
-
fusion studies confirmed that impA genes was induced by acidic
+
<hr align="center" size="2" width="100%"></span></div>
-
conditions (1),
+
<p class="MsoNormal"><a name="impA"></a><b><span
-
therefore, this is one of our candidates to complete our pH sensor
+
style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p>ImpA
-
device.</span>
+
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
<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>
style="font-size: 18pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></small></div>

Revision as of 23:50, 23 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.

 

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