http://2012.igem.org/wiki/index.php?title=Special:Contributions/Mjacobs8891&feed=atom&limit=50&target=Mjacobs8891&year=&month=2012.igem.org - User contributions [en]2024-03-28T11:17:16ZFrom 2012.igem.orgMediaWiki 1.16.0http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-04T01:37:50Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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<center><h2>Quorum STOPPING</h2></center><br />
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&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-04T01:37:08Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
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&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-04T01:36:29Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
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<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-04T01:35:42Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
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<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-04T01:35:11Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
</center><br />
<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
</center><br />
<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
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<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-04T01:31:06Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
</center><br />
<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-04T01:29:59Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
<br><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg" width=290 height=193 /><br />
</center><br />
<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/TeamTeam:CU-Boulder/Team2012-10-04T01:25:38Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<h1> The Team </h1><br />
<html><br />
<br />
<center> <img class="alignleft size-full wp-image-222" src="https://static.igem.org/mediawiki/2012/4/42/CU-Boulder_team.png" alt="Team Photo" width="525" height="350"/> <img src= "https://static.igem.org/mediawiki/2012/c/c0/CU-Boulder_logo.png" height="350"/> </center><br />
<br />
<br><br />
<a name="theplayers"></a><br />
<h2> The Players</h2> <br />
<br><br />
<br />
<table border = "1"><br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2012/4/44/45430_1453639855913_1080780129_31155161_1479673_n2.jpg" alt="Max Jacobs" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;"/> </td><td>Name: Max "Party At The Moon Tower" Jacobs<br>Majors: Molecular, Cellular, and Developmental Biology & Film Studies<br>Profession: Loch Ness Monster Enthusiast <br>iGEM focus: Inducible promoters of all kinds </td><br />
</tr><br />
<tr><br />
<td><br />
<img src=<br />
"https://static.igem.org/mediawiki/2012/f/f7/SeanIgem.jpg" alt="Sean Kalra" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;"/> </td><td> They call me Sean. I like my resuspended cells shaken, not stirred, and if you ever meet me you had better hope not to meet the things that are coming after me. </td><br />
</tr><br />
<tr><br />
<td><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/4/4c/537473_347519585318685_16517333_n.jpg" alt="Jake Sheppard" width="150" height="200" style="float:;margin:10px 10px 10px 10px;"/> </td><td> Once while sailing around the world, He discovered a short cut.<br><br />
Panhandlers give him money.<br><br />
He does Calculus in his head.<br><br />
Athletes seek his autograph.<br><br />
His passport requires no photo.<br><br />
He's never found a penny that wasn't heads up.<br><br />
When he drives his new car off the lot it increases in value.<br><br />
His 1913 Duesenberg still has that new car smell.<br><br />
Though he can't walk on water he's never slipped on ice.<br><br />
Dolphins love swimming with him. </td><br />
</tr><br />
<tr><br />
<td><br />
<img src=<br />
"https://static.igem.org/mediawiki/2012/8/8a/600670_351297528274224_907484894_n.jpg" alt="Aubrey Jackson" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;"/> </td><td> David Andrew J. "Andy" Samberg (born August 18, 1978) is an American actor, comedian, rapper, writer, and member of the comedy group The Lonely Island. He is known as a cast member on Saturday Night Live (appearing from 2005 to 2012), where he and The Lonely Island have been credited with popularizing the Emmy-winning SNL Digital Shorts, the comical short films and music videos starring Samberg and other members of the SNL cast.[2] As a film actor, Samberg has starred in Hot Rod and That's My Boy, and appeared in Space Chimps; Nick and Norah's Infinite Playlist; I Love You, Man; Cloudy with a Chance of Meatballs; Friends with Benefits; and What's Your Number?. </td><br />
</tr><br />
<tr><br />
<td><br />
<br />
<p><img src="https://static.igem.org/mediawiki/2012/b/b3/546094_2261397338944_2016106029_n.jpg" alt="Simon Greenberg" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;"/> </td><td> My name is Patrick Bateman. I am twenty-six years old. I live in the American Garden Buildings on West Eighty-First Street, on the eleventh floor.I believe in taking care of myself, in a balanced diet, in a rigorous exercise routine. In the morning, if my face is a little puffy, I'll put on an ice pack while doing my stomach crunches. I can do a thousand now. After I remove the icepack, I use a deep pore-cleanser lotion. In the shower, I use a water-activated gel cleanser, then a honey-almond body scrub, and on the face an exfoliating gel scrub.Then I apply an herb mint facial masque which I leave on for ten minutes while I prepare the rest of my routine. I always use an after-shave lotion with little or no alcohol because alcohol dries your face out and makes you look older. Then moisturizer, then an anti-aging eye balm, followed by a final moisturizing "protective" lotion...There is an idea of a Patrick Bateman, some kind of abstraction, hut there is no real me, only an <br />
entity, something illusory, and though I can hide my cold gaze and you can shake my hand and feel flesh gripping you <br />
and maybe you can even sense our lifestyles are probably comparable: I simply am not there.</td><br />
</tr><br />
</table><br />
<br><br />
<br />
<a name="theadvisors"></a><br />
<h3> The Advisors</h3><br />
<hr><br />
<br><br />
<table border = "1"><br />
<br />
<tr><td><img src="https://static.igem.org/mediawiki/2012/e/ec/Joe_picture.jpg" alt="Joe Rokicki" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;"/> </td> <td> Joe is a man about town. Child actor and molecular biology phenom, he is one of our graduate advisers. </td></tr><br />
<br />
<tr><td><img src="https://static.igem.org/mediawiki/2012/3/36/Read_t.jpg" alt="Tim Read" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;" /> </td><td> Tim is a Boston man. He enjoys candy of all forms. He is one of our graduate advisers.</td></tr><br />
<br />
<tr><td><img src="https://static.igem.org/mediawiki/2012/f/f1/Yin.jpg" alt="Hubert Yin" width="150" height="150" style="float:left;margin:10px 10px 10px 10px;"/> </td> <td> Hubert Yin is one of our faculty advisors and we work on his lab benches. </td></tr><br />
<br />
<tr><td><img src="https://static.igem.org/mediawiki/2012/d/d9/Dowell.jpg" alt="Robin Dowell" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;"/> </td> <td> Robin Dowell is another faculty advisor and we use some of her lab equipment and gel imaging computer. </td></tr><br />
<br />
<tr><td><img src="https://static.igem.org/mediawiki/2012/c/c6/Kaar.jpg" alt="Joel Kaar" width="150" height="200" style="float:left;margin:10px 10px 10px 10px;"/> </td> <td> Joel Kaar is a faculty advisor who helps us come up with concrete ideas. </td></tr><br />
<br />
</table><br />
<br />
<a name="thepics"></a><br />
<h4>The Pics </h4><br />
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<h5>Contact Us</h5><br />
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<b>Max Jacobs</b> mjacobs8891@gmail.com<br><br />
<b>Aubrey Jackson</b> aubrey.g.jackson@gmail.com<br><br />
<b>Jake Sheppard </b>sheppajm@gmail.com<br><br />
<b>Simon Greenberg </b>ajax.greenberg@gmail.com <br><br />
<b>Sean Kalra </b>nosweat444@gmail.com <br><br />
<b>Tim Read </b>timjread04@gmail.com <br><br />
<b>Joe Rokicki</b> raafekie@gmail.com<br><br />
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</html></div>Mjacobs8891http://2012.igem.org/File:Synthetic_clip_image002.jpegFile:Synthetic clip image002.jpeg2012-10-03T07:16:19Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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<br><br />
<center><h2>Quorum STOPPING</h2></center><br />
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&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="https://2012.igem.org/Team:CU-Boulder/Project">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<center><object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object></center><br />
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
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</center><br />
<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our <a href="http://www.w3schools.com/">Project</a> page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<object width="420" height="315"><param name="movie" value="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/YJWKWYQfSi0?version=3&amp;hl=en_US" type="application/x-shockwave-flash" width="420" height="315" allowscriptaccess="always" allowfullscreen="true"></embed></object><br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-03T06:51:44Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-03T06:50:47Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br><br><br />
<iframe width="420" height="315" src="http://www.youtube.com/embed/YJWKWYQfSi0" frameborder="0" allowfullscreen></iframe><br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-03T06:45:21Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br><br />
<br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-03T06:44:06Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br><br />
<p style="text-align: center><img src="https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg></p><br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-03T06:43:27Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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<hr><br />
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<p style="text-align: center>;"https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg</p><br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-03T06:43:10Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br><br><br />
<p style="text-align: center;"https://static.igem.org/mediawiki/2012/d/d1/Media_5a7257dc5595cb7be3df7954509f3e90.jpeg</p><br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence, while living in the gut of bobtail squid (shown above). By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/File:Media_5a7257dc5595cb7be3df7954509f3e90.jpegFile:Media 5a7257dc5595cb7be3df7954509f3e90.jpeg2012-10-03T06:40:57Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
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<p style="text-align: center;">https://static.igem.org/mediawiki/2012/9/94/Lux_construct.png</p><br />
<br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence. By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-03T06:37:01Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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&nbsp;&nbsp;&nbsp;&nbsp;Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Quorum sensing dictates many functions within bacteria such as when pathogenic gene products are released, biofilm formation, and food rot. Inhibiting quorum sensing became the goal of CU-Boulder’s 2012 iGEM team. This led us to find or create many interesting BioBrick parts such as Aiia, a protein that degrades specific quorums, Sdia, a transcription factor that is more sensitive to AHLs than the standard LuxR, and NucB, a nuclease that degrades preexisting biofilm. <br><br />
&nbsp;&nbsp;&nbsp;&nbsp;Instead of using a pathogenic bacteria, our team utilized the Lux gene brick from Vibrio fischeri. This gram-negative bacteria uses quorum sensing to produce bioluminescence. By using V. fischeri, our team eliminated the risk of working with pathogenic bacteria and bioluminescence proved to be easily quantifiable using plate reader experiments. As none of the presubmitted Lux bricks worked for our team, we isolated each of the 5 essential genes (LuxA,B,C,D,and E) and well an extremely important bioluminescent gene, LuxG, from a V. fischeri strain to create our model system. For more detailed information on our project visit our Projects page. <br />
<br><br><br />
The video below is an animated overview of quorum sensing.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-02T21:19:43Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br />
<table width="100%" cellpadding="0" cellspacing="10" border="0"><br />
<tr><br />
<td width="50%" valign="top"><br />
<br />
<div align=center> <b> Bacterial Nightlight </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/9b/Nemo.jpg"> </div><br />
<br />
<p>Certain organisms use bioluminescence for communication, mating, or intimidating predators. <br />
Our project aims to harness the bioluminescence genes, known as the Lux operon, from Aliivibrio fischeri,<br />
to make a genetically engineered E. Coli machine. The six genes of the 9-kilobase Lux operon, LuxCDABE and G, <br />
are regulated by LuxI and LuxR and produce blue-green light through the following reaction: </p><br />
<br />
<p style="text-align: center;"><br />
<br />
<b> FMNH2+O2+R-CHO → FMN + R-COOH + H2O + Light </b></p><br />
<br />
<p>Instead of simply transforming an E.Coli with the Lux operon under a single promoter, we have isolated <br />
each of the six genes independently and placed them under different promoters to optimize the system<br />
and produce the maximum amount of light. The entire system is regulated by a light-repressed promoter<br />
so the bacteria only luminesce during nighttime. In conclusion, we have created the first ever bacteria night light! <br />
Possible real world applications for this are endless and include illuminating street signs, marking hiking trails, and <br />
even giving light to impoverished areas of the world that do not have access to electricity.</p><br />
</td><br />
<td width="50%" valign="top"><br />
<div align=center> <b> No More Biofilm! </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/92/Mr-clean.jpg" width="306" height="208"> </div><br />
<br />
<br />
<p>Our project harnesses the use of quorum sensing to prevent bacteria from activating pathogenic factors triggered by AHLs. We developed a model system with Allivibrio fisheri. A. fisheri luminesces when activated by AHLs, which is a much safer alternative to pathogenic bacteria. We grew up our E. coli with Vibrio fisheri, and showed that the recombinant E. coli could detect V. fisheri’s AHLs, and inhibit V. fisheri from activating its quorum response genes by not producing light.The detection system then secretes factors that reduce the concentration of AHLs in the environment, thereby inhibiting the bacteria that use AHLs from communicating with each other. This is useful in preventing pathology from bacteria. When a threshold level of AHLs is reached in pathogenic bacteria, the signals activate transcription of pathogenic factors such as biofilm formation and host invasion signals (Raffa et al, 2005). We propose that disrupting the AHLs will inhibit these bacteria from creating biofilms or activating their pathogenic genes. In order to detect AHLs, we took advantage of the Salmonella enterica serovar typhimurium AHL receptor SdiA, a LuxR homolog. The SdiA transcription factor has been shown to be more sensitive to a greater diversity of AHLs than LuxR (Smith et al, 2008). Therefore it will be activated by AHLs present before the AHLs can activate genes in the surrounding bacteria. We also synthesized LuxR mutants that were more sensitive than LuxR at detecting the AHL 3OC6HSL, so they would be activated before LuxR gets activated. We planned to couple the new LuxR homologs to the AHLase Aiia, to demonstrate how we could inhibit activation of pathological factors. We also designed a construct with the new part, NucB, a nuclease that has been shown to digest extracellular DNA. </p><br />
</td><br />
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<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-02T21:19:08Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br />
<table width="100%" cellpadding="0" cellspacing="10" border="0"><br />
<tr><br />
<td width="50%" valign="top"><br />
<br />
<div align=center> <b> Bacterial Nightlight </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/9b/Nemo.jpg"> </div><br />
<br />
<p>Certain organisms use bioluminescence for communication, mating, or intimidating predators. <br />
Our project aims to harness the bioluminescence genes, known as the Lux operon, from Aliivibrio fischeri,<br />
to make a genetically engineered E. Coli machine. The six genes of the 9-kilobase Lux operon, LuxCDABE and G, <br />
are regulated by LuxI and LuxR and produce blue-green light through the following reaction: </p><br />
<br />
<p style="text-align: center;"><br />
<br />
<b> FMNH2+O2+R-CHO → FMN + R-COOH + H2O + Light </b></p><br />
<br />
<p>Instead of simply transforming an E.Coli with the Lux operon under a single promoter, we have isolated <br />
each of the six genes independently and placed them under different promoters to optimize the system<br />
and produce the maximum amount of light. The entire system is regulated by a light-repressed promoter<br />
so the bacteria only luminesce during nighttime. In conclusion, we have created the first ever bacteria night light! <br />
Possible real world applications for this are endless and include illuminating street signs, marking hiking trails, and <br />
even giving light to impoverished areas of the world that do not have access to electricity.</p><br />
</td><br />
<td width="50%" valign="top"><br />
<div align=center> <b> No More Biofilm! </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/92/Mr-clean.jpg" width="306" height="208"> </div><br />
<br />
<br />
<p>Our project harnesses the use of quorum sensing to prevent bacteria from activating pathogenic factors triggered by AHLs. We developed a model system with Allivibrio fisheri. A. fisheri luminesces when activated by AHLs, which is a much safer alternative to pathogenic bacteria. We grew up our E. coli with Vibrio fisheri, and showed that the recombinant E. coli could detect V. fisheri’s AHLs, and inhibit V. fisheri from activating its quorum response genes by not producing light.The detection system then secretes factors that reduce the concentration of AHLs in the environment, thereby inhibiting the bacteria that use AHLs from communicating with each other. This is useful in preventing pathology from bacteria. When a threshold level of AHLs is reached in pathogenic bacteria, the signals activate transcription of pathogenic factors such as biofilm formation and host invasion signals (Raffa et al, 2005). We propose that disrupting the AHLs will inhibit these bacteria from creating biofilms or activating their pathogenic genes. In order to detect AHLs, we took advantage of the Salmonella enterica serovar typhimurium AHL receptor SdiA, a LuxR homolog. The SdiA transcription factor has been shown to be more sensitive to a greater diversity of AHLs than LuxR (Smith et al, 2008). Therefore it will be activated by AHLs present before the AHLs can activate genes in the surrounding bacteria. We also synthesized LuxR mutants that were more sensitive than LuxR at detecting the AHL 3OC6HSL, so they would be activated before LuxR gets activated. We planned to couple the new LuxR homologs to the AHLase Aiia, to demonstrate how we could inhibit activation of pathological factors. We also designed a construct with the new part, NucB, a nuclease that has been shown to digest extracellular DNA. </p><br />
</td><br />
</tr><br />
</table><br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-02T21:18:13Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br />
<table width="100%" cellpadding="0" cellspacing="10" border="0"><br />
<tr><br />
<td width="50%" valign="top"><br />
<br />
<div align=center> <b> Bacterial Nightlight </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/9b/Nemo.jpg"> </div><br />
<br />
<p>Certain organisms use bioluminescence for communication, mating, or intimidating predators. <br />
Our project aims to harness the bioluminescence genes, known as the Lux operon, from Aliivibrio fischeri,<br />
to make a genetically engineered E. Coli machine. The six genes of the 9-kilobase Lux operon, LuxCDABE and G, <br />
are regulated by LuxI and LuxR and produce blue-green light through the following reaction: </p><br />
<br />
<p style="text-align: center;"><br />
<br />
<b> FMNH2+O2+R-CHO → FMN + R-COOH + H2O + Light </b></p><br />
<br />
<p>Instead of simply transforming an E.Coli with the Lux operon under a single promoter, we have isolated <br />
each of the six genes independently and placed them under different promoters to optimize the system<br />
and produce the maximum amount of light. The entire system is regulated by a light-repressed promoter<br />
so the bacteria only luminesce during nighttime. In conclusion, we have created the first ever bacteria night light! <br />
Possible real world applications for this are endless and include illuminating street signs, marking hiking trails, and <br />
even giving light to impoverished areas of the world that do not have access to electricity.</p><br />
</td><br />
<td width="50%" valign="top"><br />
<div align=center> <b> No More Biofilm! </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/92/Mr-clean.jpg" width="306" height="208"> </div><br />
<br />
<br />
<p>Our project harnesses the use of quorum sensing to prevent bacteria from activating pathogenic factors triggered by AHLs. We developed a model system with Allivibrio fisheri. A. fisheri luminesces when activated by AHLs, which is a much safer alternative to pathogenic bacteria. We grew up our E. coli with Vibrio fisheri, and showed that the recombinant E. coli could detect V. fisheri’s AHLs, and inhibit V. fisheri from activating its quorum response genes by not producing light.The detection system then secretes factors that reduce the concentration of AHLs in the environment, thereby inhibiting the bacteria that use AHLs from communicating with each other. This is useful in preventing pathology from bacteria. When a threshold level of AHLs is reached in pathogenic bacteria, the signals activate transcription of pathogenic factors such as biofilm formation and host invasion signals (Raffa et al, 2005). We propose that disrupting the AHLs will inhibit these bacteria from creating biofilms or activating their pathogenic genes. In order to detect AHLs, we took advantage of the Salmonella enterica serovar typhimurium AHL receptor SdiA, a LuxR homolog. The SdiA transcription factor has been shown to be more sensitive to a greater diversity of AHLs than LuxR (Smith et al, 2008). Therefore it will be activated by AHLs present before the AHLs can activate genes in the surrounding bacteria. We also synthesized LuxR mutants that were more sensitive than LuxR at detecting the AHL 3OC6HSL, so they would be activated before LuxR gets activated. We planned to couple the new LuxR homologs to the AHLase Aiia, to demonstrate how we could inhibit activation of pathological factors. We also designed a construct with the new part, NucB, a nuclease that has been shown to digest extracellular DNA. </p><br />
</td><br />
</tr><br />
</table><br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-02T21:16:52Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br />
<table width="100%" cellpadding="0" cellspacing="10" border="0"><br />
<tr><br />
<td width="50%" valign="top"><br />
<br />
<div align=center> <b> Bacterial Nightlight </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/9b/Nemo.jpg"> </div><br />
<br />
<p>Certain organisms use bioluminescence for communication, mating, or intimidating predators. <br />
Our project aims to harness the bioluminescence genes, known as the Lux operon, from Aliivibrio fischeri,<br />
to make a genetically engineered E. Coli machine. The six genes of the 9-kilobase Lux operon, LuxCDABE and G, <br />
are regulated by LuxI and LuxR and produce blue-green light through the following reaction: </p><br />
<br />
<p style="text-align: center;"><br />
<br />
<b> FMNH2+O2+R-CHO → FMN + R-COOH + H2O + Light </b></p><br />
<br />
<p>Instead of simply transforming an E.Coli with the Lux operon under a single promoter, we have isolated <br />
each of the six genes independently and placed them under different promoters to optimize the system<br />
and produce the maximum amount of light. The entire system is regulated by a light-repressed promoter<br />
so the bacteria only luminesce during nighttime. In conclusion, we have created the first ever bacteria night light! <br />
Possible real world applications for this are endless and include illuminating street signs, marking hiking trails, and <br />
even giving light to impoverished areas of the world that do not have access to electricity.</p><br />
</td><br />
<td width="50%" valign="top"><br />
<div align=center> <b> No More Biofilm! </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/92/Mr-clean.jpg" width="306" height="208"> </div><br />
<br />
<br />
<p>Our project harnesses the use of quorum sensing to prevent bacteria from activating pathogenic factors triggered by AHLs. We developed a model system with Allivibrio fisheri. A. fisheri luminesces when activated by AHLs, which is a much safer alternative to pathogenic bacteria. We grew up our E. coli with Vibrio fisheri, and showed that the recombinant E. coli could detect V. fisheri’s AHLs, and inhibit V. fisheri from activating its quorum response genes by not producing light.The detection system then secretes factors that reduce the concentration of AHLs in the environment, thereby inhibiting the bacteria that use AHLs from communicating with each other. This is useful in preventing pathology from bacteria. When a threshold level of AHLs is reached in pathogenic bacteria, the signals activate transcription of pathogenic factors such as biofilm formation and host invasion signals (Raffa et al, 2005). We propose that disrupting the AHLs will inhibit these bacteria from creating biofilms or activating their pathogenic genes. In order to detect AHLs, we took advantage of the Salmonella enterica serovar typhimurium AHL receptor SdiA, a LuxR homolog. The SdiA transcription factor has been shown to be more sensitive to a greater diversity of AHLs than LuxR (Smith et al, 2008). Therefore it will be activated by AHLs present before the AHLs can activate genes in the surrounding bacteria. We also synthesized LuxR mutants that were more sensitive than LuxR at detecting the AHL 3OC6HSL, so they would be activated before LuxR gets activated. We planned to couple the new LuxR homologs to the AHLase Aiia, to demonstrate how we could inhibit activation of pathological factors. We also designed a construct with the new part, NucB, a nuclease that has been shown to digest extracellular DNA. </p><br />
</td><br />
</tr><br />
</table><br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-10-02T21:15:58Z<p>Mjacobs8891: </p>
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<br><br />
<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
<br />
<hr><br />
<br />
<table width="100%" cellpadding="0" cellspacing="10" border="0"><br />
<tr><br />
<td width="50%" valign="top"><br />
<br />
<div align=center> <b> Bacterial Nightlight </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/9b/Nemo.jpg"> </div><br />
<br />
<p>Certain organisms use bioluminescence for communication, mating, or intimidating predators. <br />
Our project aims to harness the bioluminescence genes, known as the Lux operon, from Aliivibrio fischeri,<br />
to make a genetically engineered E. Coli machine. The six genes of the 9-kilobase Lux operon, LuxCDABE and G, <br />
are regulated by LuxI and LuxR and produce blue-green light through the following reaction: </p><br />
<br />
<p style="text-align: center;"><br />
<br />
<b> FMNH2+O2+R-CHO → FMN + R-COOH + H2O + Light </b></p><br />
<br />
<p>Instead of simply transforming an E.Coli with the Lux operon under a single promoter, we have isolated <br />
each of the six genes independently and placed them under different promoters to optimize the system<br />
and produce the maximum amount of light. The entire system is regulated by a light-repressed promoter<br />
so the bacteria only luminesce during nighttime. In conclusion, we have created the first ever bacteria night light! <br />
Possible real world applications for this are endless and include illuminating street signs, marking hiking trails, and <br />
even giving light to impoverished areas of the world that do not have access to electricity.</p><br />
</td><br />
<td width="50%" valign="top"><br />
<div align=center> <b> No More Biofilm! </b> </div> <br><br />
<br />
<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/92/Mr-clean.jpg" width="306" height="208"> </div><br />
<br />
<br />
<p>Our project harnesses the use of quorum sensing to prevent bacteria from activating pathogenic factors triggered by AHLs. We developed a model system with Allivibrio fisheri. A. fisheri luminesces when activated by AHLs, which is a much safer alternative to pathogenic bacteria. We grew up our E. coli with Vibrio fisheri, and showed that the recombinant E. coli could detect V. fisheri’s AHLs, and inhibit V. fisheri from activating its quorum response genes by not producing light.The detection system then secretes factors that reduce the concentration of AHLs in the environment, thereby inhibiting the bacteria that use AHLs from communicating with each other. This is useful in preventing pathology from bacteria. When a threshold level of AHLs is reached in pathogenic bacteria, the signals activate transcription of pathogenic factors such as biofilm formation and host invasion signals (Raffa et al, 2005). We propose that disrupting the AHLs will inhibit these bacteria from creating biofilms or activating their pathogenic genes. In order to detect AHLs, we took advantage of the Salmonella enterica serovar typhimurium AHL receptor SdiA, a LuxR homolog. The SdiA transcription factor has been shown to be more sensitive to a greater diversity of AHLs than LuxR (Smith et al, 2008). Therefore it will be activated by AHLs present before the AHLs can activate genes in the surrounding bacteria. We also synthesized LuxR mutants that were more sensitive than LuxR at detecting the AHL 3OC6HSL, so they would be activated before LuxR gets activated. We planned to couple the new LuxR homologs to the AHLase Aiia, to demonstrate how we could inhibit activation of pathological factors. We also designed a construct with the new part, NucB, a nuclease that has been shown to digest extracellular DNA. </p><br />
</td><br />
</tr><br />
</table><br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/File:Biofront2.jpgFile:Biofront2.jpg2012-10-02T15:43:29Z<p>Mjacobs8891: </p>
<hr />
<div></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/ProjectTeam:CU-Boulder/Project2012-10-02T15:06:22Z<p>Mjacobs8891: </p>
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<h1><p style="text-align: center;">Project 1: Model System for studying quorum activation of pathogenicity</p></h1><br />
<br><br><br><br />
Quorum sensing is a system of bacterial communication that coordinates gene expression based on population density. Bacteria secrete a signaling molecule, such as N-Acyl Homoserine Lactones (AHL), and each bacteria has a extracellular receptor that specifically binds the signaling molecule which activates transcription factors specific to certain genes. Many pathogenic bacteria use quorum sensing to activate their virulent factors, as it is not energetically favorable for a single bacterium to release its virulent factors until it is in a large enough population for infect a host. For example, Pseudomonas Aeruginosa is an opportunistic pathogen that contributes to sepsis and its virulent proteins are not expressed constitutively but in a density dependent manner.(1)(2). In our project the CU-Boulder iGEM team aims to disrupt quorum sensing in order to reduce pathogenicity of otherwise virulent bacteria.<br><br />
It has been well documented in research that bacteria have used quorum sensing signals to activate pathogenic factors. The gram negative bacteria P. aeruginosa synthesizes AHLs as a bacterial density indicator, and upon reaching a specific threshold, pathogenic factors are activated (Iglewski et al. 2000). P. aeruginosa is a common opportunistic infection that occurs most frequently in immunocompromised people (Iglewski et al. 2000). Though this same quorum sensing mechanism for activation of pathogenic traits has also been demonstrated in Salmonella, E. coli, and Y. pestis.(-----). Quorum sensing factors also play a role in activating multi-bacterial responses such as biofilm production, and food rot. (…) In order to study how to inhibit these pathogenic effects of quorum sensing, we needed a system that would respond to quorum signals, and was a safe alternative to the pathogenic bacteria. Therefore, we characterized the model quorum sensing system of Allivibrio fisheri, which luminesces in response to AHLs.<br />
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Allivibrio fischeri is a gram-negative bacteria that is a biological model for both quorum-sensing and bioluminescence research. This rod-shaped bacteria participates in symbiotic relationships with various marine organisms, most notably the Bobtail squid. This squid hosts Allivibrio fisheri and uses its light to hide the squid's shadow in shallow waters to avoid potential predators. We decided to use Allivibrio fisheri WH1 strain as our model organism because it is a safe organism to handle, it has a LuxI/LuxR quorum sensing system that it usees to detect the concentration of AHLs, and it responds with luminescence at a visible wavelength when the threshold level of AHLs is reached.<br />
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When LuxR detects AHLs it activates the rest of the Lux operon. The Lux operon (below) consists of 5 essential genes: LuxA, LuxB, LuxC, LuxD, and LuxE.<br><br />
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<p style="text-align: center;">https://static.igem.org/mediawiki/2012/a/a4/Fig9.png</p><br />
LuxC, LuxD, and Lux E form the fatty acid reductase enzyme complex that synthesizes fatty acid aldehydes with acyl-CoA as the starting substrate. The 3 gene products form a complex conglomerate (below) to increase kinetic efficiency. LuxD is the transferase, LuxE is the synthetase, and LuxC is the reductase. <br />
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<p style="text-align: center;">https://static.igem.org/mediawiki/2012/9/94/Lux_construct.png</p><br />
<br>LuxA and LuxB form a dimer that reduces O<sub>2</sub> while oxidizing the aldehyde and FMNH<sub>2</sub>. This oxidation produces the blue-green light (~490nm). Another representation of the overall reaction is shown below:<br><br />
<p style="text-align: center;">https://static.igem.org/mediawiki/2012/4/42/Lux_reaction.png</p><br />
Also, it has been shown that the LuxG gene, although not essential, does increase brightness by functioning as a flavin reductase.(http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2258676/)<br />
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We utilized Allivibrio fisheri because the luminosity is easy to measure with a high degree of sensitivity and low background noise. The organism is visually luminous at a wavelength that can be perceived by the human eye, and we can therefore use the organism as a model for reporting when pathogenic factors would have been initiated in other bacteria. Using a plate reader, we characterized at what OD the A. fisheri starts to glow, and characterized how the density of the organism affected the luminosity. (Figure)<br />
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'''Basic Idea:''' Transform an E. Coli with the 5 essential Lux genes (LuxABCDE) as well as Lux G. This Lux brick will be under the control of a UV induced promoter system and a C1 lambda inverter system. This means that while the bacteria is exposed to UV light, the C1 protein will repress C1 regulated promoter that is upstream of the Lux brick, and no light will be produced. On the flip side, when the bacteria is not exposed to UV light, no C1 will be made and the C1 regulated promoter will be activated and produce the Lux brick and the culture will luminesce. To further optimize the system, we have expressed the individual Lux genes in varying levels to see which of the 6 is the rate limiting step in bioluminescence. <br />
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'''Possible Promoter systems:'''<br />
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Circadian clock - KaiABC/SasA/RpaA is the simplest oscillator found in cyanobacteria, Synechococcus elongatus<br />
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Blue Light Induced - YcgE/YcgF is endogenous to E. Coli. We have the system built and are testing it now.<br />
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Red light Induced - Cph8/PCB/OmpR is a system that won iGEM in 2006 but every team that has used it since has run into problems<br />
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<h3><p style="text-align: center;">Project 1: Experiments</p></h3><br />
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1. '''Assemble the LuxABCDE operon + LuxG''' The Cambridge 2010 "E. Glowi" team submitted the Lux operon to the registry, BBa_K325905, but several attempts at transforming E. Coli with this part failed either due to its large size (6478bp) or due to an inaccurate registry submission. Several other teams have tried to use their part since 2010 but none have been successful. For this reason, our team is BioBricking each Lux gene individually and submitting them to the registry.<br><br><br />
2. '''Optimize the Lux System''' Once the Lux brick is working in a cell, we ligated each Lux gene to a different strength promoter to maximize luminescence while conserving energy. Because LuxC is the scaffolding on which the LuxCDE reductase complex is built, we started by up regulating LuxC under a high constitutive promoter.<br><br><br />
3. '''Regulate the system under a UV induced promoter''' We ligated a UV induced promoter to a C1 lambda inverter system which is upstream of the optimized Lux Brick. This is our final project as the Lux Brick will only luminesce in low light conditions (night) and the system will be repressed in high light conditions (daytime). Visit our Modeling page to see how we used rapidly degrading LVA tags to ensure a quickly responding system.<br><br />
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<h3><p style="text-align: center;">Project 1: Results</p></h3><br />
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<h1><p style="text-align: center;">Project 2: Biofilm Prevention</p></h1><br />
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'''Quorum Sensing in LuxR containing Bacteria<br />
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This year the CU iGEM team has harnessed the quorum sensing factors endogenous in the Salmonella enterica serovar typhimurium LT2 strain to detect AHLs produced by other bacteria. Neither Salmonella nor E. coli have been shown to produce detectable levels of AHLs, so they were model organisms for the detection of other AHL producing bacteria such as Vibrio fisheri, or pathogenic bacteria such as Yersinia pestis. AHLs are small molecules synthesized by most gram negative bacteria, and are able to freely diffuse throughout the membrane. When the concentration of AHL producing bacteria in a specific area increases, the total concentration of AHLs diffusing into the cytoplasm also increases. Once a threshold level of AHLs are in the cytosol, transcription factors like the Vibrio fisheri LuxR or Salmonella enterica SdiA activate gene synthesis for quorum factors to be produced. In pathogenic bacteria, these signals have been shown to activate transcription of pathogenic factors.<br />
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[[File:Slide1.jpg]]<br />
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'''Detection and silencing of AHL producing bacteria'''<br />
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The SdiA transcription factor has been shown to be more sensitive to lower concentrations and a greater diversity of AHLs than its LuxR homolog. We took advantage of this extra-sensitivity to create a detection system for AHL producing bacteria, while additionally attaching the detection system to the secretion of potent AHLases such as the Aiia enzyme, and a reporter protein RFP. Using this construct we tested whether we could disrupt the quorum sensing signal AHLs in order to keep them from reaching the threshold concentration to activate the less sensitive LuxR receptor. The V. fisheri species and other pathogenic bacteria use the LuxR receptor, therefore inhibition of AHLs would keep V. fisheri from producing light. Keeping the V. fisheri from producing light is an analogous model to keeping pathogenic bacteria from being stimulated by AHLs to release their pathogenesis factors. <br />
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[[File:Slide2.jpg]]<br />
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Inhibiting quorum sensing in bacteria has been shown to not only inhibit the transcription of pathogenesis factors, but also to keep biofilms from forming. In the history of iGEM teams have used secreted enzymes to digest biofilms that have already been made, but our use of quorum sensing inhibitors have gone a step further, and are being tested in their ability to keep bacteria from making biofilms in the first place.<br />
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<h3><p style="text-align: center;">Project 2: Experiments</p></h3><br />
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1. '''Test whether the SdiA pSrgE cassette is more sensitive to extracellular AHLs than the LuxR LuxpR cassette.'''<br />
This will be done by placing an RFP behind the cassette, and using the plate reader to determine the fluorescence at a given concentration of RFP.<br />
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2. '''Use the SdiA pSrgE cassette’s sensitivity to inhibit the LuxR LuxpR system'''. <br />
Attach an AHLase and YFP to the more sensitive SdiA/pSrgE cassette. Co-culture the AHLase/YFP construct with a LuxR/LuxpR-RFP bacteria. The culture should turn fluoresce yellow, and not turn red.<br />
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3. '''Test the SdiA pSrgE cassette’s inhibition effects with Vibrio fisheri acting as a pathogen analogue.'''<br />
Co-culture the SdiA/pSrgE/YFP/Aiia cells with V. fisheri and use the plate reader to show that co-cultures will keep V. fisheri from emitting light at given ODs. <br />
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<h3><p style="text-align: center;">Project 2: Results</p></h3><br />
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<h1><p style="text-align: center;">Applications</p></h1><br />
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<h1><p style="text-align: center;">Medal Consideration</p></h1><br />
<h3><p style="text-align: left;">Bronze</p></h3><br />
<ul><br />
<p style="text-align: left;">https://static.igem.org/mediawiki/2012/7/79/Small-checkmark.png Team Registration:<i> April 11, 2012</i> </p><br />
<li>___ Complete Judging form</li><br />
<p style="text-align: left;">https://static.igem.org/mediawiki/2012/7/79/Small-checkmark.png Team Wiki</p><br />
<li> ___ Present Poster and a talk at the iGEM Jamboree:<i> North American West Regional Jamboree at Stanford University on Oct.16, 2012</i></li><br />
<li>___ At least one new submitted BioBrick part or a new application of an existing part with quantitative documentation</li><br />
</ul><br />
<h3><p style="text-align: left;">Silver</p></h3><br />
<ul><br />
<li> ___ Demonstrate that a newly submitted BioBrick part works as expected</li><br />
<li> ___ Characterize the operation of at least one new BioBrick Part and enter this information in the Registry</li><br />
</ul><br />
<h3><p style="text-align: left;">Gold</p></h3><br />
<p style="text-align: left;">'''Complete one or more of the following'''</p><br />
<ul><br />
<li> ___ Improve the function of an existing BioBrick part and enter it's information in the registry</li><br />
<li> ___ Help another iGEM team by, for example, characterizing a part, debugging a construct, or modeling or simulating their system</li><br />
<li> ___ Outline and detail a new approach to an issue of Human Practice in synthetic biology as it relates to your project, such as safety, security, ethics, or ownership, sharing, and innovation</li><br />
</ul></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:42:35Z<p>Mjacobs8891: </p>
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<center><h2>Safety</h2></center><br />
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<h5>1. Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
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Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
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Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
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Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
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<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No. The parts produced by our group do not raise any safety issues.<br />
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<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found <a href="http://www.colorado.edu/ehs/research/biological.html"> here</a><br />
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<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
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Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"/"target="_blank"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:23:28Z<p>Mjacobs8891: </p>
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<div>{{CU-Boulder-Header}}<br />
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<center><h2>Safety</h2></center><br />
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<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
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Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No. The parts produced by our group do not raise any safety issues.<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found <a href="http://www.colorado.edu/ehs/research/biological.html"> here</a><br />
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<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
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Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"/"target="_blank"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:22:48Z<p>Mjacobs8891: </p>
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<div>{{CU-Boulder-Header}}<br />
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<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No. The parts produced by our group have all been promoter BioBrick parts and they do not raise any safety issues.<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found <a href="http://www.colorado.edu/ehs/research/biological.html"> here</a><br />
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<br><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"/"target="_blank"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/AttributionsTeam:CU-Boulder/Attributions2012-09-07T23:20:49Z<p>Mjacobs8891: </p>
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<h1><p style="text-align: center;">A Special Thanks to our Primary Sponsors!</p></h1><br />
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<p style="text-align: center;"> Click on a logo to open their homepage in a new window</p><br />
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<p style="text-align: center;"><a href="http://cimb.colorado.edu/"target="_blank"><img src="https://static.igem.org/mediawiki/2012/9/91/IQBiology.jpeg" height="143" width="604"></a><br />
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<p style="text-align: center;"><a href="http://enrichment.colorado.edu/urop/"target="_blank"><img src="https://static.igem.org/mediawiki/2012/c/c0/UROP_logo.png" height="79" width="810"></a><br />
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<p style="text-align: center;"><a href="http://www.goldlabcolorado.com/"target="_blank"><img src="https://static.igem.org/mediawiki/2012/7/73/GoldLab_logo.jpg" height="148" width="678" ></a><br />
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<h2><p style="text-align: center;">We Would Also Like to Thank our Product Sponsors</p></h2><br />
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<td><a href="http://www.neb.com/nebecomm/default.asp"target="_blank"><img src="https://static.igem.org/mediawiki/2012/6/65/NEW_ENGLAND_BIOLABS.PNG " height="88" width="221" border="0"/></td><br />
<td><a href="http://www.promega.com/"target="_blank"><img src="https://static.igem.org/mediawiki/2012/c/c4/Promega.jpg" height="112" width="184" border="0"/> </a><br />
<td><a href="http://www.vwr.com/"target="_blank"><img src="https://static.igem.org/mediawiki/2012/4/42/VWR_logo.jpg" height="72" width="244" border="0"/></td></tr><br />
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<td><a href="http://www.lucigen.com/"target="_blank"><img src="https://static.igem.org/mediawiki/2012/a/a9/Lucigen-logo.jpg" border="0" /></a></td><br />
<td><a href="http://www.idtdna.com/site"target="_blank"><img src="https://static.igem.org/mediawiki/2012/2/22/IDT-Logo.jpg" height="120" width="289" border="0"/> </a><br />
</td><br />
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</html></div>Mjacobs8891http://2012.igem.org/File:IQBiology.jpegFile:IQBiology.jpeg2012-09-07T23:18:56Z<p>Mjacobs8891: </p>
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<div></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:15:55Z<p>Mjacobs8891: </p>
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<div>{{CU-Boulder-Header}}<br />
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<br />
<html><br />
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<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No. The parts produced by our group have all been promoter BioBrick parts and they do not raise any safety issues.<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found <a href="http://www.colorado.edu/ehs/research/biological.html"> here</a><br />
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<br><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"/"target="_blank"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:10:52Z<p>Mjacobs8891: </p>
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<div>{{CU-Boulder-Header}}<br />
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<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No. The parts produced by our group have all been promoter BioBrick parts and they do not raise any safety issues.<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found <a href="http://www.colorado.edu/ehs/research/biological.html"> here</a><br />
<br><br />
<br> <br />
<br><br />
<br><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:09:06Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found <a href="http://www.colorado.edu/ehs/research/biological.html"> here</a><br />
<br><br />
<br> <br />
<br><br />
<br><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:08:35Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found <a href="http://www.colorado.edu/ehs/research/biological.html."> here</a><br />
<br><br />
<br> <br />
<br><br />
<br><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:07:30Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found at <a href="http://www.colorado.edu/ehs/research/biological.html."></a><br />
<br><br />
<br> <br />
<br><br />
<br><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:06:58Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found at <ahref="http://www.colorado.edu/ehs/research/biological.html."></a><br />
<br><br />
<br> <br />
<br><br />
<br><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T23:06:28Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No<br />
<br><br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h5><br />
The CU-Boulder iGEM team complied with standard Institutional Biosafety Commitee requirements. This includes but is not limited to proper decontamination procedures, sharps disposed of properly, gloves worn, working with approved agents and protocols, waste properly decontaminated, disposed, packaged, labeled, no eating, drinking, contact lenses or food storage in the lab. More details about the Institutional Biosafety Committee safety standards can be found at <a href="http://www.colorado.edu/ehs/research/biological.html."></a><br />
<br><br />
<br> <br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T22:57:12Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5> No<br />
<br><br />
<h5> 3. Is there a local biosafety group, committee, or review board at your institution?</h><br />
<br><br />
<br> <br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T22:55:43Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5> 2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5><br />
<br><br />
<br><br />
<br> <br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T22:55:28Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<center><h2>Safety</h2></center><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<h5>Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h5><br />
<br><br />
<br><br />
<br> <br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T22:52:07Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<h2>Safety</h2><br />
<br />
<h5>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h5> <br><br />
<br><br />
An AHLase, Aiia, and nuclease, NucB, are incorporated into an E. coli construct, which acts to disrupt biofilm formation and degrade any present biofilm. This is a proof of concept that could eventually be taken into plants or used as a probiotic. <br><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<br><br />
<br><br />
<br> <br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T22:49:47Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
<h3>1) Would any of your project ideas raise safety issues in terms of researcher, environmental, and public safety?</h3> <br><br />
<br><br />
An AHLase, Aiia, and nuclease, NucB, are incorporated into an E. coli construct, which acts to disrupt biofilm formation and degrade any present biofilm. This is a proof of concept that could eventually be taken into plants or used as a probiotic. <br><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<br><br />
<br><br />
<br> <br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
<br />
</html></div>Mjacobs8891http://2012.igem.org/Team:CU-Boulder/SafetyTeam:CU-Boulder/Safety2012-09-07T22:44:06Z<p>Mjacobs8891: </p>
<hr />
<div>{{CU-Boulder-Header}}<br />
<br />
<br />
<br />
<html><br />
<br />
1) An AHLase, Aiia, and nuclease, NucB, are incorporated into an E. coli construct, which acts to disrupt biofilm formation and degrade any present biofilm. This is a proof of concept that could eventually be taken into plants or used as a probiotic. <br><br />
<br><br />
<br />
Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br><br />
<br><br />
<br />
Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br><br />
<br><br />
<br />
Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame.<br><br />
<br><br />
<br><br />
<br> <br />
<br />
<center><img src="https://static.igem.org/mediawiki/2012/6/6b/Unicycle.jpeg" height="360" width="560"><br />
<br><br />
Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
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1) An AHLase, Aiia, and nuclease, NucB, are incorporated into an E. coli construct, which acts to disrupt biofilm formation and degrade any present biofilm. This is a proof of concept that could eventually be taken into plants or used as a probiotic. <br />
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Researcher safety: In its current state, the construct does not pose a threat or raise safety issues for the researchers working on it. All work with E. coli was conducted using standard laboratory protocol and organic waste was disposed of in common biohazard procedure. DH5-alpha E. coli cells were used for all transformations (both chemically competent and electrically competent).<br />
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Public safety: No modifications were made to the E. coli that would alter its pathogenicity to humans. DH5-alpha cells were used, which pose no risk of disease that could be spread to the public. The idea of the eventual use as a probiotic would need to be strictly tested in a controlled manner in the future if the construct proved to be effective. For the IGEM project, the construct was solely a proof of concept.<br />
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Environmental safety: Similarly to public safety, if the construct was to be tested in plants, it would need to be done in a safe manner under experimental conditions. Considering the scope of this project, the construct was not transformed into any form of plant cell, as that would require much more advanced safety procedures and a longer time frame. <br />
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Use this page to answer the questions on the <a href="https://2012.igem.org/Team:CU-Boulder/Safety"> safety page </a>.<br />
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</html></div>Mjacobs8891http://2012.igem.org/Team:CU-BoulderTeam:CU-Boulder2012-07-09T17:43:01Z<p>Mjacobs8891: </p>
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<p style="text-align: center;">Welcome to the CU-iGEM 2012 wiki! This is the first time the University of Colorado has competed in the International Genetically Engineered Machines competition, and we are excited to show you the science coming out of Boulder, Colorado! We are a team of five undergraduates advised by our two graduate mentors, and a smattering of post-docs. This year is also the grand opening of the brand new Biofrontiers building for bioscience, and biotechnology research. We have been fortunate enough to have lab space in the new building.</p><br />
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<div align=center> <b> Bacterial Nightlight </b> </div> <br><br />
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<div align=center> <img src="https://static.igem.org/mediawiki/2012/9/9b/Nemo.jpg"> </div><br />
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<p>Certain organisms use bioluminescence for communication, mating, or intimidating predators. <br />
Our project aims to harness the bioluminescence genes, known as the Lux operon, from Aliivibrio fischeri,<br />
to make a genetically engineered E. Coli machine. The six genes of the 9-kilobase Lux operon, LuxCDABE and G, <br />
are regulated by LuxI and LuxR and produce blue-green light through the following reaction: </p><br />
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<b> FMNH2+O2+R-CHO → FMN + R-COOH + H2O + Light </b></p><br />
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<p>Instead of simply transforming an E.Coli with the Lux operon under a single promoter, we have isolated <br />
each of the six genes independently and placed them under different promoters to optimize the system<br />
and produce the maximum amount of light. The entire system is regulated by a light-repressed promoter<br />
so the bacteria only luminesce during nighttime. In conclusion, we have created the first ever bacteria night light! <br />
Possible real world applications for this are endless and include illuminating street signs, marking hiking trails, and <br />
even giving light to impoverished areas of the world that do not have access to electricity.</p><br />
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<div align=center> <b> No More Biofilm! </b> </div> <br><br />
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<p>Our second project harnesses the use of quorum sensing to detect bacteria that secrete AHLs. The detection system then secretes factors that reduce the concentration of AHLs in the environment, thereby confusing the bacteria that use AHLs to communicate with each other. This is useful in preventing pathology from bacteria. When a threshold level of AHLs is reached in pathogenic bacteria, the signals activate transcription of pathogenic factors such as biofilm formation and host invasion signals. Disrupting the AHLs will inhibit these bacteria from creating biofilms or activating their pathogenic genes. We took advantage of the Salmonella enterica serovar typhimurium AHL receptor SdiA, a LuxR homolog. The SdiA transcription factor has been shown to be more sensitive to lower concentrations and a greater diversity of AHLs than LuxR. Therefore it will be activated by AHLs present before the AHLs can activate genes in the surrounding bacteria. We coupled it with both an RFP and an AHLase, Aiia. As a proof of concept, we grew up our E. coli with Vibrio fisheri, and showed that the recombinant E. coli could detect V. fisheri’s AHLs, and inhibit V. fisheri from activating its quorum response genes by not producing light. </p><br />
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