Team:UC Davis/Adding secretion/model 1

From 2009.igem.org

(Difference between revisions)

Revision as of 07:12, 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 148: / Line 148: @@
 are intending to harness and make use of this feature by fusing a
 specific
-protein to it. <o:p></o:p></span></p>
+protein to it. <o:p></o:p></span><br>
-<p class="MsoNormal" style="line-height: normal;"><i><span
+<i><span
 style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
 have
-modified this protein to Biobrick standard, Tom Knights Standard.</span></i><span
+modified this protein to Biobrick standard, Tom Knights Standard.<br>
+For more information go to: <a
+href="http://partsregistry.org/wiki/index.php/Part:BBa_K265008">BBa_K265008</a>&nbsp;</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"
@@ Line 177: / Line 179: @@
 have
 modified this protein to Biobrick standard, Tom Knights Standard.</span></i><span
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span></p>
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span><br>
-<p class="MsoNormal" style="line-height: normal;"><i><span
+<i><span
 style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Note:
 “It has
@@ Line 184: / Line 186: @@
 sorted
 and incorporated into this membrane” (10)</span></i><span
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span></p>
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <o:p></o:p></span><br>
-<p class="MsoNormal" style="line-height: normal;"><i><span
+<i><span
 style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
 more
-information go to:</span></i><span
+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="color: blue;">http://partsregistry.org/wiki/index.php?title=Part:BBa_J36836</span></i></a><o:p></o:p></span></p>
+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;"
@@ Line 203: / Line 207: @@
 style="font-size: 12pt; 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. <o:p></o:p></span></p>
+from the registry is being used in our secretion system. <o:p></o:p></span><br>
-<p class="MsoNormal" style="line-height: normal;"><i><span
+<i><span
 style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
 more
 information go to:</span></i><span
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <a
+style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <big><a
+href="http://partsregistry.org/wiki/index.php/Part:BBa_J61132"><small><span
+style="font-style: italic;">BBa_J61132</span></small></a></big><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="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;"
@@ Line 221: / Line 227: @@
 style="font-size: 12pt; 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.<o:p></o:p></span></p>
+our terminator in both our secretion and pH system.<o:p></o:p></span><br>
-<p class="MsoNormal" style="line-height: normal;"><i><span
+<i><span
 style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">For
 more
 information go to:</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_B0015"><span
+style="font-style: italic;">BBa_B0015</span></a> <a
 href="http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015"><i><span
-style="color: blue;">http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015</span></i></a><o:p></o:p></span></p>
+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;"
@@ Line 240: / Line 248: @@
 are using Green Fluorescent Protein as a
 reporter that also serves as a small protein in testing our secretion
-system.<o:p></o:p></span></p>
+system.<br>
+<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;"
@@ Line 252: / Line 264: @@
 is a firefly protein that also
 fluoresces, so it serves as a reporter as well as a testable large
-protein. <o:p></o:p></span></p>
+protein.<br>
+<span style="font-style: italic;">For more information go to: <a
+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;"
@@ Line 265: / Line 280: @@
 part registry.<br>
 </span><i><span
-style="font-size: 13pt; 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;;"></span></i><i><span
-</span></i><i><span
+style="font-size: 12pt; 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;;">More
+more information go to:<a
-can be
+href="http://partsregistry.org/wiki/index.php/Part:BBa_R0010">
-found in: </span></i><span
+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="color: blue;">http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010</span></i></a><o:p></o:p></span></p>
+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;"
@@ Line 285: / Line 301: @@
 INPNC, contains a cleavable site that allows the target fusion protein
 to
-‘secrete’ from INPNC. We will do the same with OmpA. <o:p></o:p></span></p>
+‘secrete’ from INPNC. We will do the same with OmpA. <o:p></o:p></span><br>
-<p class="MsoNormal" style="line-height: normal;"><i><span
+<i><span
 style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">We
 have
-modified this protein to Biobrick standard, Tom Knights Standard.</span></i><span
+modified this protein to Biobrick standard, Tom Knights Standard.<br>
+For more infromation go to :<a
+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"
@@ Line 296: / Line 314: @@
 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="line-height: normal;"><a name="His"></a><b><span
+<a name="His"></a><b><span
 style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">6-His
 Tag</span></b><span
@@ Line 304: / Line 322: @@
 Blotting if our fluorescent reporters are not expressed as highly as we
 would
-like. <o:p></o:p></span></p>
+like. <o:p></o:p></span><br>
-<p class="MsoNormal" style="line-height: normal;"><i><span
+<small><i><span
 style="font-size: 12pt; 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
+plate reader.</span></i></small>
-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;"