Team:UC Davis/Adding secretion/model 4

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Revision as of 07:16, 2 October 2009

Model 1

=

Secretion Model 1:

Click on an individual part for more information.

a.

b.

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). 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.
For more information go to: BBa_K265008

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 E. coli outer membrane are sorted and incorporated into this membrane” (10)
For more information go to: BBa_K103006

RBS: Ribosome Binding site number 32 (BBa_J61132) from the registry is being used in our secretion system.
For more information go to: 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: BBa_B0015

GFP: We are using Green Fluorescent Protein as a reporter that also serves as a small protein in testing our secretion system.
For more informaiton go to: BBa_K265003

Luciferase: Luciferase is a firefly protein that also fluoresces, so it serves as a reporter as well as a testable large protein.
For more information go to: BBa_1712019

LacI: One inducible Promoter which was found in the part registry.
For more information go to: 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.
For more infromation go to :BBa_K265002

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.

@@ Line 132: / Line 132: @@
 <big><big><big><span
 style="text-decoration: underline; font-weight: bold;"></span></big></big></big>
-<hr style="width: 100%; height: 2px;">
+<hr style="width: 100%; height: 2px;"><b><span
-<p class="MsoNormal" style=""><b><span
+style="font-size: 13.5pt; 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;;"><a
+name="INPNC"></a>INPNC:</span></b><span
-name="INPNC"></a>INPNC</span></b><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Ice-nucleation
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:
+protein (INP) from Pseudomonas
-Ice-nucleation protein (INP) from Pseudomonas Syringae was
+Syringae was suggested to be used for display of foreign proteins on
-suggested to be used for display of foreign proteins on the surface of <i>E.coli</i>(7).Furthermore,
+the
-researches have shown that an INP derivative constituting the N-and
+surface of <i>E. coli</i>(7).Furthermore, researches have shown that
-C-terminal
+an INP
-domains can and has been used to display foreign proteins on the
+derivative constituting the N-and C-terminal domains can and has been
-surface of <i>E.coli</i>(9).
+used to
-</span><span style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+display foreign proteins on the surface of <i>E. coli</i>(9). In our
-<p class="MsoNormal" style=""><i><span
+project we
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
+are intending to harness and make use of this feature by fusing a
-A study has shown that "Ice- nucleation Protein (INP), an outer
+specific
-membrane
+protein to it. <o:p></o:p></span><br>
-protein from Pseudomonas syringae, is able to catalyze the ice crystal
+<p class="MsoNormal" style="line-height: normal;"><i><span
-formation of supercooled water.</span></i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">In
+have
-our project we are intending to harness and make use of this feature by
+modified this protein to Biobrick standard, Tom Knights Standard.<br>
-fusing
+For more information go to: <a
-a specific protein to it. <i>We have modified this protein to Biobrick
+href="http://partsregistry.org/wiki/index.php/Part:BBa_K265008">BBa_K265008</a>&nbsp;</span></i><span
-standard, Tom Knights Standard.</i></span><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span></p>
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+<div class="MsoNormal"
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+align="center"><span
+style="font-size: 12pt; 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
+<p class="MsoNormal" style="line-height: normal;"><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; 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;;">:
+style="font-size: 18pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:</span><span
-OmpA is one of the proteins on the outer membrane of <i>E.coli</i>
+style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">&nbsp;</span><span
-(13). OmpA has been found to be useful as utilizable fusion part that
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">OmpA
-can fuse
+is one of the proteins on the outer
-our protein to and display on the surface of <i>E.coli</i>. This part
+membrane of <i>E. coli</i> (13). OmpA has been found to be useful as
-has
+utilizable
-already been documented on the parts registry; however, it has not been
+fusion part that can fuse our protein to and display on the surface of <i>E.
-tested
+coli</i>. This part has already been documented on the parts registry;
-via fusion with a target protein linked with a cleavable signal
+however,
-sequence.<u1:p></u1:p></span><span
+it has not been tested via fusion with a target protein linked with a
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
+cleavable
-</span><i><span
+signal sequence. <o:p></o:p></span></p>
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
+<p class="MsoNormal" style="line-height: normal;"><i><span
-have modified this protein to Biobrick standard, Tom Knights
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
-Standard. </span></i><span
+have
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><u1:p></u1:p><o:p></o:p></span></p>
+modified this protein to Biobrick standard, Tom Knights Standard.</span></i><span
-<p class="MsoNormal" style=""><i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span><br>
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:</span></i><span
+<i><span
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
-“<i>It has
+“It has
-remained essentially unknown how proteins of Escherichia coli outer
+remained essentially unknown how proteins of E. coli outer membrane are
-membrane
+sorted
-are sorted and incorporated into this membrane” (10)<br>
+and incorporated into this membrane” (10)</span></i><span
-<u1:p></u1:p>For more information go to:</i> <a
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span><br>
+<i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
+more
+information go to:<a
+href="http://partsregistry.org/wiki/index.php/Part:BBa_K103006">
+BBa_K103006</a></span></i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <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="color: blue;"></span></i></a><o:p></o:p></span></p>
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+<div class="MsoNormal"
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+align="center"><span
+style="font-size: 12pt; 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
+<p class="MsoNormal" style="line-height: normal;"><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; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">RBS</span></b><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;;">:&nbsp;
-Ribosome Binding site number 32 (BBa_J61132) from the registry is
+</span><span
-being used in our secretion system.<u1:p></u1:p></span><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Ribosome
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
+Binding site number 32 (BBa_J61132)
-</span><i><span
+from the registry is being used in our secretion system. <o:p></o:p></span><br>
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
+<i><span
-more information go to:</span></i><span
+style="font-size: 12pt; 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
-<a href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><i><span
+information go to:</span></i><span
-style="">http://partsregistry.org/wiki/index.php/Part:BBa_J61132</span></i></a></span><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <big><a
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><small><span
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+style="font-style: italic;">BBa_J61132</span></small></a></big><a
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><i><span
+style="color: blue;"></span></i></a><o:p></o:p></span></p>
+<div class="MsoNormal"
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
+align="center"><span
+style="font-size: 12pt; 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
+<p class="MsoNormal" style="line-height: normal;"><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; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Terminator</span></b><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;;">: </span><span
-We are using BBa_B0015, a double terminator, as our terminator in
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
-both our secretion and pH system.<u1:p></u1:p></span><span
+are using BBa_B0015, a double terminator, as
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
+our terminator in both our secretion and pH system.<o:p></o:p></span><br>
-</span><i><span
+<i><span
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
-more information go to:</span></i><span
+more
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+information go to:</span></i><span
-<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015"><i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <a
-style="">http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015</span></i></a></span><span
+href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015"><span
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+style="font-style: italic;">BBa_B0015</span></a> <a
-<div class="MsoNormal" style="text-align: center;" align="center"><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;;">
+style="color: blue;"></span></i></a><o:p></o:p></span></p>
+<div class="MsoNormal"
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
+align="center"><span
+style="font-size: 12pt; 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
+<p class="MsoNormal" style="line-height: normal;"><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; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">GFP</span></b><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;;">: </span><span
-We are using Green Fluorescent Protein as a reporter that also
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
-serves as a small protein in testing our secretion system.</span><span
+are using Green Fluorescent Protein as a
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+reporter that also serves as a small protein in testing our secretion
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+system.<br>
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+<span style="font-style: italic;">For more informaiton go to: </span><a
+style="font-style: italic;"
+href="http://partsregistry.org/wiki/index.php/Part:BBa_K265003">BBa_K265003</a><span
+style="font-style: italic;">&nbsp;</span><o:p></o:p></span></p>
+<div class="MsoNormal"
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
+align="center"><span
+style="font-size: 12pt; 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
+<p class="MsoNormal" style="line-height: normal;"><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; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Luciferase</span></b><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;;">: </span><span
-Luciferase is a firefly protein that also fluoresces, so it
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Luciferase
-serves as a reporter as well as a testable large protein.<i><br>
+is a firefly protein that also
-More can be found in:</i></span><span
+fluoresces, so it serves as a reporter as well as a testable large
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+protein.<br>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+<span style="font-style: italic;">For more information go to: <a
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+href="http://partsregistry.org/wiki/index.php/Part:BBa_I712019">BBa_1712019</a></span>
+<o:p></o:p></span></p>
+<div class="MsoNormal"
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
+align="center"><span
+style="font-size: 12pt; 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
+<p class="MsoNormal" style="line-height: normal;"><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; 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;;">:
+style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">: </span><span
-One inducible Promoter which was found in the part registry.<i><br>
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">One
-More can be found in: </i><a
+inducible Promoter which was found in the
+part registry.<br>
+</span><i><span
+style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></i><i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
+more information go to:<a
+href="http://partsregistry.org/wiki/index.php/Part:BBa_R0010">
+BBa_R0010</a><br>
+</span></i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><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="color: blue;"></span></i></a><o:p></o:p></span></p>
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+<div class="MsoNormal"
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+align="center"><span
+style="font-size: 12pt; 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
+<p class="MsoNormal" style="line-height: normal;"><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; 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;;">:
+style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:</span><span
-This signal sequence, when placed between INPNC, contains a
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">This
-cleavable site that allows the target fusion protein to ‘secrete’ from
+signal sequence, when placed between
-INPNC.&nbsp; We will do the same with OmpA.&nbsp; <u1:p></u1:p></span><span
+INPNC, contains a cleavable site that allows the target fusion protein
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+to
-<p class="MsoNormal" style=""><i><span
+‘secrete’ from INPNC. We will do the same with OmpA. <o:p></o:p></span><br>
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
+<i><span
-have modified this protein to Biobrick standard, Tom Knights
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
-Standard.</span></i><span
+have
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+modified this protein to Biobrick standard, Tom Knights Standard.<br>
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+For more infromation go to :<a
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+href="http://partsregistry.org/wiki/index.php/Part:BBa_K265002">BBa_K265002</a></span></i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span></p>
+<div class="MsoNormal"
+style="margin-bottom: 0.0001pt; text-align: center; line-height: normal;"
+align="center"><span
+style="font-size: 12pt; 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
+<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
+style="font-size: 13pt; 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;;">:
+style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">:</span><span
-The 6-Histidine Tag serves as a tag for Western Blotting if our
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">The
-fluorescent reporters are not expressed as highly as we would like.<u1:p></u1:p></span><span
+-Histidine Tag serves as a tag for Western
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
+Blotting if our fluorescent reporters are not expressed as highly as we
-</span><i><span
+would
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
+like. <o:p></o:p></span><br>
-We are using this tag, just in case if the GFP or Luciferase
+<small><i><span
-does not work under a plate reader.</span></i><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></p>
+We are
-<div class="MsoNormal" style="text-align: center;" align="center"><span
+using this tag, just in case if the GFP or Luciferase does not work
-style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">
+under a
-<hr align="center" size="2" width="100%"></span></div>
+plate reader.<br>
-<p class="MsoNormal" style=""><a name="ChvI_promoter"></a><b><span
+</span></i></small>
-style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">ChvI
+<hr style="width: 100%; height: 2px;"><small><i><span
-promoter</span></b><span
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></i></small>
-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.<br>
-</span></p>
-<hr style="width: 100%; height: 2px;">
 <p class="MsoNormal"><br>
 <span