http://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&feed=atom&action=historyTeam:Cambridge/Bioluminescence/G28 - Revision history2024-03-29T11:59:39ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=199810&oldid=prevAnjaFides at 23:02, 27 October 20102010-10-27T23:02:48Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/RightImage|image=Space_invader.jpg|caption=The LuxBrick on a 96 well plate}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/RightImage|image=Space_invader.jpg|caption=The LuxBrick on a 96 well plate}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of ''Vibrio fischeri'''] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete ''V. fischeri'' lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in ''V. fischeri''. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of ''Vibrio fischeri'''] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete ''V. fischeri'' lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in ''V. fischeri''. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td></tr>
</table>AnjaFideshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=199774&oldid=prevAnjaFides at 23:02, 27 October 20102010-10-27T23:02:04Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/headerbar|colour=#386abc|caption=Project Vibrio: The LuxBrick}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/headerbar|colour=#386abc|caption=Project Vibrio: The LuxBrick}}</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/RightImage|image=Space_invader.jpg|caption=The LuxBrick on a 96 well plate}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/RightImage|image=Space_invader.jpg|caption=The LuxBrick on a 96 well plate}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of ''Vibrio fischeri'''] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete ''V. fischeri'' lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in ''V. fischeri''. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of ''Vibrio fischeri'''] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete ''V. fischeri'' lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in ''V. fischeri''. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td></tr>
</table>AnjaFideshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=191059&oldid=prevTheos at 18:13, 27 October 20102010-10-27T18:13:13Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/RightImage|image=Jungle_book.jpg|caption<del class="diffchange diffchange-inline">:</del>Illuminating the Jungle Book with the LuxBrick}}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/RightImage|image=Jungle_book.jpg|caption<ins class="diffchange diffchange-inline">=</ins>Illuminating the Jungle Book with the LuxBrick}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </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>
</table>Theoshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=191046&oldid=prevTheos at 18:12, 27 October 20102010-10-27T18:12:49Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/headerMinimalprototype}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/headerMinimalprototype}}</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/headerbar|colour=#386abc|<del class="diffchange diffchange-inline">title</del>=Project Vibrio: The LuxBrick}}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/headerbar|colour=#386abc|<ins class="diffchange diffchange-inline">caption</ins>=Project Vibrio: The LuxBrick}}</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">{{:Team:Cambridge/Templates/rightpic|src=Jungle_book.jpg|300px|right|G28 illuminating the Jungle Book}}</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/<del class="diffchange diffchange-inline">rightpic</del>|<del class="diffchange diffchange-inline">src</del>=Space_invader.jpg|<del class="diffchange diffchange-inline">400px|right|G28 </del>on a 96 well plate}}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/<ins class="diffchange diffchange-inline">RightImage</ins>|<ins class="diffchange diffchange-inline">image</ins>=Space_invader.jpg|<ins class="diffchange diffchange-inline">caption=The LuxBrick </ins>on a 96 well plate}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of ''Vibrio fischeri'''] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete ''V. fischeri'' lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in ''V. fischeri''. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of ''Vibrio fischeri'''] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete ''V. fischeri'' lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in ''V. fischeri''. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><div style="clear:both"></div></ins></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 colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">{{:Team:Cambridge/Templates/RightImage|image=Jungle_book.jpg|caption:Illuminating the Jungle Book with the LuxBrick}}</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </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>
</table>Theoshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=171654&oldid=prevTheos: /* h-ns mutants */2010-10-27T02:24:37Z<p><span class="autocomment">h-ns mutants</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>To test the theory that H-NS proteins repress the Lux genes by interacting with their coding regions, we transformed two E.coli mutant strains with this construct and measured their light output. The strains we used were GM230 hns-205::Tn10, which has a C-terminal deletion in the H-NS gene and BW25113 Δhns::kan, a strain from a knockout library. In the literature, h-ns mutant strains have been described as producing much brighter luminescence than wild type strains. While we could not reproduce a higher peak brightness, it was apparent that the knockout strain maintained its light output for much longer than wild type, which showed a steep reduction in brightness upon entering stationary phase. This phenomenon (Abrupt Decline in Luciferase Activity - or ADLA) has been described by [http://www.springerlink.com/content/w73k840k27866462/fulltext.pdf Koga et al. 2004]. However, this paper suggests that the effect of H-NS on luminescence is not due to expression of the lux genes, but occurs indirectly via the cell's redox pool, in particular the availability of FMNH2.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>To test the theory that H-NS proteins repress the Lux genes by interacting with their coding regions, we transformed two E.coli mutant strains with this construct and measured their light output. The strains we used were GM230 hns-205::Tn10, which has a C-terminal deletion in the H-NS gene and BW25113 Δhns::kan, a strain from a knockout library. In the literature, h-ns mutant strains have been described as producing much brighter luminescence than wild type strains. </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>While we could not reproduce a higher peak brightness, it was apparent that the knockout strain maintained its light output for much longer than wild type, which showed a steep reduction in brightness upon entering stationary phase. This phenomenon (Abrupt Decline in Luciferase Activity - or ADLA) has been described by [http://www.springerlink.com/content/w73k840k27866462/fulltext.pdf Koga et al. 2004]. However, this paper suggests that the effect of H-NS on luminescence is not due to expression of the lux genes, but occurs indirectly via the cell's redox pool, in particular the availability of FMNH2.</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: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/footerMinimal}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/footerMinimal}}</div></td></tr>
</table>Theoshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=166660&oldid=prevAnjaFides at 21:15, 26 October 20102010-10-26T21:15:26Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of Vibrio fischeri'] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete V. fischeri lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in V. fischeri. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of <ins class="diffchange diffchange-inline">''</ins>Vibrio fischeri<ins class="diffchange diffchange-inline">''</ins>'] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete <ins class="diffchange diffchange-inline">''</ins>V. fischeri<ins class="diffchange diffchange-inline">'' </ins>lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in <ins class="diffchange diffchange-inline">''</ins>V. fischeri<ins class="diffchange diffchange-inline">''</ins>. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</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>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting V. fischeri) under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting <ins class="diffchange diffchange-inline">''</ins>V. fischeri<ins class="diffchange diffchange-inline">''</ins>) under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </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: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td></tr>
</table>AnjaFideshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=166621&oldid=prevAnjaFides at 21:12, 26 October 20102010-10-26T21:12:22Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Jungle_book.jpg|300px|right|G28 illuminating the Jungle Book}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Jungle_book.jpg|300px|right|G28 illuminating the Jungle Book}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of Vibrio fischeri <del class="diffchange diffchange-inline">[...]</del>'] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete V. fischeri lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in V. fischeri. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of Vibrio fischeri'] by James Slock from King's College, PA. Dr Slock kindly provided us the two plasmids mentioned in these experiments, which carry the complete V. fischeri lux operon (LuxICDABE regulated by Lux R). Using Long-Range PCR, we extracted luxCD, luxAB and luxE individually and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], lux I and lux R exert tight quorum sensing control on expression of the lux operon in V. fischeri. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</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: #eee; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting V. fischeri) under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to generate an operon consisting of Lux C, D, A, B, E (in this order, reflecting V. fischeri) under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td></tr>
</table>AnjaFideshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=166610&oldid=prevAnjaFides at 21:11, 26 October 20102010-10-26T21:11:55Z<p></p>
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<col class='diff-marker' />
<col class='diff-content' />
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 21:11, 26 October 2010</td>
</tr><tr><td colspan="2" class="diff-lineno">Line 3:</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Jungle_book.jpg|300px|right|G28 illuminating the Jungle Book}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Jungle_book.jpg|300px|right|G28 illuminating the Jungle Book}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>James Slock from King's College, PA kindly provided us <del class="diffchange diffchange-inline">with plasmids carrying </del>the <del class="diffchange diffchange-inline">genes responsible for bioluminescence </del>in <del class="diffchange diffchange-inline">''</del>V. fischeri<del class="diffchange diffchange-inline">''</del>. Using Long-Range PCR, we extracted <del class="diffchange diffchange-inline">these genes </del>and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], the lux operon in <del class="diffchange diffchange-inline">''</del>V. fischeri<del class="diffchange diffchange-inline">'' is under tight quorum sensing control</del>. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Whilst researching the literature on bioluminescence at the beginning of the summer, we came across a report on [http://departments.kings.edu/biology/lux/bacterial.html 'Molecular Biology Experiments Utilizing the lux Genes of Vibrio fischeri [...]'] by </ins>James Slock from King's College, PA<ins class="diffchange diffchange-inline">. Dr Slock </ins>kindly provided us the <ins class="diffchange diffchange-inline">two plasmids mentioned </ins>in <ins class="diffchange diffchange-inline">these experiments, which carry the complete </ins>V. fischeri <ins class="diffchange diffchange-inline">lux operon (LuxICDABE regulated by Lux R)</ins>. Using Long-Range PCR, we extracted <ins class="diffchange diffchange-inline">luxCD, luxAB and luxE individually </ins>and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], <ins class="diffchange diffchange-inline">lux I and lux R exert tight quorum sensing control on expression of </ins>the lux operon in V. fischeri. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</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>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to <del class="diffchange diffchange-inline">produce </del>an operon consisting of Lux C, D, A, B, E (in this order, reflecting <del class="diffchange diffchange-inline">''</del>V. fischeri<del class="diffchange diffchange-inline">''</del>) under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to <ins class="diffchange diffchange-inline">generate </ins>an operon consisting of Lux C, D, A, B, E <ins class="diffchange diffchange-inline"> </ins>(in this order, reflecting V. fischeri) under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct the LuxBrick. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </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: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td></tr>
</table>AnjaFideshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=159747&oldid=prevTheos at 13:34, 26 October 20102010-10-26T13:34:43Z<p></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 13:34, 26 October 2010</td>
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<td colspan="2" class="diff-lineno">Line 5:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>James Slock from King's College, PA kindly provided us with plasmids carrying the genes responsible for bioluminescence in ''V. fischeri''. Using Long-Range PCR, we extracted these genes and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], the lux operon in ''V. fischeri'' is under tight quorum sensing control. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>James Slock from King's College, PA kindly provided us with plasmids carrying the genes responsible for bioluminescence in ''V. fischeri''. Using Long-Range PCR, we extracted these genes and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], the lux operon in ''V. fischeri'' is under tight quorum sensing control. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</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>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to produce an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct <del class="diffchange diffchange-inline">G28</del>. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to produce an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct <ins class="diffchange diffchange-inline">the LuxBrick</ins>. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </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: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td></tr>
</table>Theoshttp://2010.igem.org/wiki/index.php?title=Team:Cambridge/Bioluminescence/G28&diff=158322&oldid=prevTheos at 11:43, 26 October 20102010-10-26T11:43:47Z<p></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 11:43, 26 October 2010</td>
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<td colspan="2" class="diff-lineno">Line 5:</td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>James Slock from King's College, PA kindly provided us with plasmids carrying the genes responsible for bioluminescence in ''V. fischeri''. Using Long-Range PCR, we extracted these genes and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], the lux operon in ''V. fischeri'' is under tight quorum sensing control. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>James Slock from King's College, PA kindly provided us with plasmids carrying the genes responsible for bioluminescence in ''V. fischeri''. Using Long-Range PCR, we extracted these genes and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], the lux operon in ''V. fischeri'' is under tight quorum sensing control. In the absence of LuxR protein and AHL the Lux genes are virtually inactive.</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"> </del>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to produce an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct G28. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to produce an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct G28. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part. </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: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=h-ns mutants=</div></td></tr>
</table>Theos