http://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&feed=atom&action=historyTeam:Penn State/Project - Revision history2024-03-29T06:40:20ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=206390&oldid=prevLer172: /* Overall project */2010-10-28T03:00:25Z<p><span class="autocomment">Overall project</span></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>This system could also be implemented in other chassis, such as yeast (wine) or Lactobacillus (yogurt), so the food itself could be the sensor. Genetically Modified Organisms (GMOs) in food would obviously face severe regulatory constraints. To better understand public perception, we created and administered a survey to over 250 randomized students in cooperation with other iGEM teams, especially RMIT. For details, see our [[Human Practices|Human Practices]] page.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>This system could also be implemented in other chassis, such as yeast (wine) or Lactobacillus (yogurt), so the food itself could be the sensor. Genetically Modified Organisms (GMOs) in food would obviously face severe regulatory constraints. To better understand public perception, we created and administered a survey to over 250 randomized students in cooperation with other iGEM teams, especially RMIT. For details, see our [[<ins class="diffchange diffchange-inline">Team:Penn State/</ins>Human Practices|Human Practices]] page.</div></td></tr>
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</table>Ler172http://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=206259&oldid=prevLer172: /* cI mechanism */2010-10-28T02:56:54Z<p><span class="autocomment">cI mechanism</span></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The Tet inverter behaves somewhat as a constitutive promoter when no promoter is in front of it. Following the Tet inverter is a cI coding sequence (which codes for the lambda repressor). The lambda repressor binds to the lux promoters in the front of the circuit, which are also lambda repressible. This, coupled with the fact that the conditions are anaerobic, provides a very strong "<del class="diffchange diffchange-inline">off</del>" switch that does not allow the expression of the Anaerobic Fluorescent Protein (AFP) to occur.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The Tet inverter behaves somewhat as a constitutive promoter when no promoter is in front of it. Following the Tet inverter is a cI coding sequence (which codes for the lambda repressor). The lambda repressor binds to the lux promoters in the front of the circuit, which are also lambda repressible. This, coupled with the fact that the conditions are anaerobic, provides a very strong "<ins class="diffchange diffchange-inline">OFF</ins>" switch that does not allow the expression of the Anaerobic Fluorescent Protein (AFP) to occur.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===FNR breaks apart in oxygen===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===FNR breaks apart in oxygen===</div></td></tr>
</table>Ler172http://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=205140&oldid=prevAjk5319: /* Inconsistent Sequencing Results */2010-10-28T02:15:04Z<p><span class="autocomment">Inconsistent Sequencing Results</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Certain parts on the registry have reportedly had inconsistent sequencing results, or not enough information is available to say for certain that the actual part is what its creators think it is. Below is a list of parts for which the Penn State iGEM team repeatedly received the same but inaccurate sequencing results. These findings have been posted on each part's registry page in order to hopefully start a discussion with other teams who may have experienced the same results.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Certain parts on the registry have reportedly had inconsistent sequencing results, or not enough information is available to say for certain that the actual part is what its creators think it is. Below is a list of parts for which the Penn State iGEM team repeatedly received the same but inaccurate sequencing results. These findings have been posted on each part's registry page in order to hopefully start a discussion with other teams who may have experienced the same results.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>constitutive promoter [http://partsregistry.org/Part:BBa_J23108 J23108]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>constitutive promoter [http://partsregistry.org/Part:BBa_J23108<ins class="diffchange diffchange-inline">:Experience#User_Reviews </ins>J23108]</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>RBS+LuxR [http://partsregistry.org/<del class="diffchange diffchange-inline">wiki/index.php?title=</del>Part:BBa_J37033 J37033]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>RBS+LuxR [http://partsregistry.org/Part:BBa_J37033<ins class="diffchange diffchange-inline">:Experience </ins>J37033]</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Lux promoter [http://partsregistry.org/Part:BBa_K091107 K091107]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Lux promoter [http://partsregistry.org/Part:BBa_K091107<ins class="diffchange diffchange-inline">:Experience#User_Reviews </ins>K091107]</div></td></tr>
</table>Ajk5319http://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=203880&oldid=prevAjk5319: /* Inconsistent Sequencing Results */2010-10-28T01:19:01Z<p><span class="autocomment">Inconsistent Sequencing Results</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Certain parts on the registry have reportedly had inconsistent sequencing results, or not enough information is available to say for certain that the actual part is what its creators think it is. Below is a list of parts for which the Penn State iGEM team repeatedly received the same but inaccurate sequencing results. These findings have been posted on each part's registry page in order to hopefully start a discussion with other teams who may have experienced the same results.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Certain parts on the registry have reportedly had inconsistent sequencing results, or not enough information is available to say for certain that the actual part is what its creators think it is. Below is a list of parts for which the Penn State iGEM team repeatedly received the same but inaccurate sequencing results. These findings have been posted on each part's registry page in order to hopefully start a discussion with other teams who may have experienced the same results.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>constitutive promoter <del class="diffchange diffchange-inline">[</del>[http://partsregistry.org/Part:BBa_J23108 J23108<del class="diffchange diffchange-inline">]</del>]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>constitutive promoter [http://partsregistry.org/Part:BBa_J23108 J23108]</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>RBS+LuxR <del class="diffchange diffchange-inline">[</del>[http://partsregistry.org/wiki/index.php?title=Part:BBa_J37033 J37033<del class="diffchange diffchange-inline">]</del>]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>RBS+LuxR [http://partsregistry.org/wiki/index.php?title=Part:BBa_J37033 J37033]</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Lux promoter <del class="diffchange diffchange-inline">[</del>[http://partsregistry.org/Part:BBa_K091107 K091107<del class="diffchange diffchange-inline">]</del>]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Lux promoter [http://partsregistry.org/Part:BBa_K091107 K091107]</div></td></tr>
</table>Ajk5319http://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=203813&oldid=prevAjk5319: /* Inconsistent Sequencing Results */2010-10-28T01:17:19Z<p><span class="autocomment">Inconsistent Sequencing Results</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Inconsistent Sequencing Results====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Inconsistent Sequencing Results====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">The </del>constitutive promoter [[J23108]] <del class="diffchange diffchange-inline">and </del>RBS+LuxR [[<del class="diffchange diffchange-inline">C0062</del>]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Certain parts on the registry have reportedly had inconsistent sequencing results, or not enough information is available to say for certain that the actual part is what its creators think it is. Below is a list of parts for which the Penn State iGEM team repeatedly received the same but inaccurate sequencing results. These findings have been posted on each part's registry page in order to hopefully start a discussion with other teams who may have experienced the same results.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>constitutive promoter [[<ins class="diffchange diffchange-inline">http://partsregistry.org/Part:BBa_J23108 </ins>J23108]]</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>RBS+LuxR [[<ins class="diffchange diffchange-inline">http://partsregistry.org/wiki/index.php?title=Part:BBa_J37033 J37033]]</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Lux promoter [[http://partsregistry.org/Part:BBa_K091107 K091107</ins>]]</div></td></tr>
</table>Ajk5319http://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=200891&oldid=prevErik: /* Lux Promoter Characterization */2010-10-27T23:40:04Z<p><span class="autocomment">Lux Promoter Characterization</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Two Lux-inducible promoters that were already on the registry (K091107 and K091146) were characterized. Because these promoters require AHL and LuxR in order to activate, a construct was created that consisted of a Lux promoter followed by RFP, and a constitutive promoter followed by the LuxR gene. LuxR was expected to be present in excess so that the limiting factor would be AHL. The coding sequences for LuxR and RFP were preceded by the standard RBS B0034. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Two Lux-inducible promoters that were already on the registry (K091107 and K091146) were characterized. Because these promoters require AHL and LuxR in order to activate, a construct was created that consisted of a Lux promoter followed by RFP, and a constitutive promoter followed by the LuxR gene. LuxR was expected to be present in excess so that the limiting factor would be AHL. The coding sequences for LuxR and RFP were preceded by the standard RBS B0034. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The AHL used was <del class="diffchange diffchange-inline">N(Beta-Ketocaproyl)-L</del>-homoserine lactone. It was added to samples in a 96-well plate in concentrations of 0, .1, 1, 10, 100 and 1000nM.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The AHL used was <ins class="diffchange diffchange-inline">OC6</ins>-homoserine lactone. It was added to samples in a 96-well plate in concentrations of 0, .1, 1, 10, 100 and 1000nM<ins class="diffchange diffchange-inline">.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The results below show no trend in protein expression based on concentration of OC6-homoserine lactone. More research is warranted to provide information about what are the best conditions for chemically inducing the Lux promoters</ins>.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:1107_108.png|800px]]</center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:1107_108.png|800px]]</center></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:146_116_tetR.png|800px]]</center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:146_116_tetR.png|800px]]</center></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">These results showed no trend in protein expression based on concentration of N(Beta-Ketocaproyl)-L-homoserine lactone. More research is warranted to provide information about what are the best conditions for chemically inducing the Lux promoters.</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">====Inconsistent Sequencing Results====</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The constitutive promoter [[J23108]] and RBS+LuxR [[C0062]]</ins></div></td></tr>
</table>Erikhttp://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=200755&oldid=prevErik: /* Characterization of Old BioBrick parts */2010-10-27T23:32:10Z<p><span class="autocomment">Characterization of Old BioBrick parts</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Characterization of Old BioBrick parts===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Characterization of Old BioBrick parts===</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>When designing our project, we were thinking about using the violacein pigment, BBa_K274002, in our genetic circuit. This part is supposed to be just the coding sequence <del class="diffchange diffchange-inline">without a </del>promoter. <del class="diffchange diffchange-inline">As </del>can be seen <del class="diffchange diffchange-inline">by the picture </del>below, the <del class="diffchange diffchange-inline">coding sequence appears to have some sort of constitutive </del>promoter <del class="diffchange diffchange-inline">attached to it</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>When designing our project, we were thinking about using the violacein pigment, BBa_K274002, in our genetic circuit. This part is supposed to be just the coding sequence<ins class="diffchange diffchange-inline">. Because the tube in the picture below contains only this part with no </ins>promoter <ins class="diffchange diffchange-inline">added, there should be no expression of violacein</ins>. <ins class="diffchange diffchange-inline">However, as </ins>can be seen below, the <ins class="diffchange diffchange-inline">violacein is present. This could be caused by either an undocumented </ins>promoter <ins class="diffchange diffchange-inline">within the part, or a non-insulated BioBrick vector with inefficient flanking transcriptional terminators</ins>.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:Violacein.jpg|400px]]</center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:Violacein.jpg|400px]]</center></div></td></tr>
</table>Erikhttp://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=200663&oldid=prevErik: /* Results */2010-10-27T23:26:26Z<p><span class="autocomment">Results</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:RawODoxygen.png|800px]]</center></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center>[[Image:RawODoxygen.png|800px]]</center></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Shown below is the Fluorescence over OD of each sample. As the samples reach steady state, the graph should become a horizontal line. Notice how the graph comes closer and closer to reaching this steady state with each successive plate. Also, it is important to <del class="diffchange diffchange-inline">rememeber </del>that evaporation comes into play here when comparing anaerobic and anaerobic samples with each other. Since the anaerobic samples are covered in mineral oil, it is expected that the evaporation rates of the media underneath is less. This means that, for aerobic cells, the OD becomes artificially inflated as the same number of cells have become concentrated in less and less media. It is not known to what extent the evaporation effect has on these data.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Shown below is the Fluorescence over OD of each sample. As the samples reach steady state, the graph should become a horizontal line. Notice how the graph comes closer and closer to reaching this steady state with each successive plate. Also, it is important to <ins class="diffchange diffchange-inline">remember </ins>that evaporation comes into play here when comparing anaerobic and anaerobic samples with each other. Since the anaerobic samples are covered in mineral oil, it is expected that the evaporation rates of the media underneath is less. This means that, for aerobic cells, the OD becomes artificially inflated as the same number of cells have become concentrated in less and less media. It is not known to what extent the evaporation effect has on these data.</div></td></tr>
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</table>Erikhttp://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=200570&oldid=prevErik: /* Oxygen Sensor */2010-10-27T23:22:58Z<p><span class="autocomment">Oxygen Sensor</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Oxygen Sensor ===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Oxygen Sensor ===</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The design of the oxygen-sensing promoter was based on the dcuC gene of E. coli. The two binding sites for FNR surrounded the RNAP binding site and were systematically shifted upstream and downstream from this site in multiple constructs, causing overlap of the RNAP binding site in some cases. Using this strategy, five constructs were designed with the constitutive promoter J23113 as the basis for the sequence following the downstream FNR binding site. The construct <del class="diffchange diffchange-inline">out of these five </del>that worked best as an oxygen-sensing promoter was fully characterized and is shown below.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The design of the oxygen-sensing promoter was based on the dcuC gene of E. coli. The two binding sites for FNR surrounded the RNAP binding site and were systematically shifted upstream and downstream from this site in multiple constructs, causing overlap of the RNAP binding site in some cases. Using this strategy, five constructs were designed with the constitutive promoter J23113 as the basis for the sequence following the downstream FNR binding site. The construct that worked best as an oxygen-sensing promoter <ins class="diffchange diffchange-inline">(J6, [[http://partsregistry.org/Part:BBa_K376003 BBa_J376003]]) </ins>was fully characterized and is shown below.</div></td></tr>
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</table>Erikhttp://2010.igem.org/wiki/index.php?title=Team:Penn_State/Project&diff=200389&oldid=prevErik: /* Oxygen Sensor */2010-10-27T23:18:57Z<p><span class="autocomment">Oxygen Sensor</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Oxygen Sensor ===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=== Oxygen Sensor ===</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The design of the oxygen-sensing promoter was based on the dcuC gene of E. coli. The two binding sites for FNR surrounded the RNAP binding site and were systematically shifted upstream and downstream from this site in multiple constructs, causing overlap of the RNAP binding site in some cases. <del class="diffchange diffchange-inline">Five </del>constructs were designed <del class="diffchange diffchange-inline">using </del>the constitutive promoter J23113 as the basis for the <del class="diffchange diffchange-inline">rest of the promoter </del>following the downstream FNR binding site. The construct out of these five that was fully characterized <del class="diffchange diffchange-inline">was J6, </del>and <del class="diffchange diffchange-inline">its sequence </del>is <del class="diffchange diffchange-inline">as follows:</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The design of the oxygen-sensing promoter was based on the dcuC gene of E. coli. The two binding sites for FNR surrounded the RNAP binding site and were systematically shifted upstream and downstream from this site in multiple constructs, causing overlap of the RNAP binding site in some cases. <ins class="diffchange diffchange-inline">Using this strategy, five </ins>constructs were designed <ins class="diffchange diffchange-inline">with </ins>the constitutive promoter J23113 as the basis for the <ins class="diffchange diffchange-inline">sequence </ins>following the downstream FNR binding site. The construct out of these five that <ins class="diffchange diffchange-inline">worked best as an oxygen-sensing promoter </ins>was fully characterized and is <ins class="diffchange diffchange-inline">shown below.</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The oxygen promoters were characterized using a Tecan infinite M1000 microplate reader. Each sample was grown overnight on plates and transferred to a 96 deep-well plate for overnight growth. After 16 hours the samples were tested in the Tecan, where measurements would be taken for around 12 hours. Sometimes, measurement of growth of three successive plates (serially diluted from the preceding one) was needed before the appropriate steady state could be recorded.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The oxygen promoters were characterized using a Tecan infinite M1000 microplate reader. Each sample was grown overnight on plates and transferred to a 96 deep-well plate for overnight growth. After 16 hours the samples were tested in the Tecan, where measurements would be taken for around 12 hours. Sometimes, measurement of growth of three successive plates (serially diluted from the preceding one) was needed before the appropriate steady state could be recorded.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== Results ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== Results ==</div></td></tr>
</table>Erik