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<center><h2>Quorum STOPPING</h2></center>
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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>
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&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://2012.igem.org/Team:CU-Boulder/Project">Project</a> page.
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            <div align=center> <b> Bacterial Nightlight </b> </div> <br>
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The video below is an animated overview of quorum sensing.
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<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>
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            <p>Certain organisms use bioluminescence for communication, mating, or intimidating predators.  
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            Our project aims to harness the bioluminescence genes, known as the Lux operon, from Aliivibrio fischeri,
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            to make a genetically engineered E. Coli machine. The six genes of the 9-kilobase Lux operon, LuxCDABE and G,  
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            are regulated by LuxI and LuxR and produce blue-green light through the following reaction: </p>
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                        <b> FMNH2+O2+R-CHO → FMN + R-COOH + H2O + Light </b></p>
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          <p>Instead of simply transforming an E.Coli with the Lux operon under a single promoter, we have isolated
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          each of the six genes independently and placed them under different promoters to optimize the system
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          and produce the maximum amount of light. The entire system is regulated by a light-repressed promoter
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          so the bacteria only luminesce during nighttime. In conclusion, we have created the first ever bacteria night light!
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          Possible real world applications for this are endless and include illuminating street signs, marking hiking trails, and  
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          even giving light to impoverished areas of the world that do not have access to electricity.</p>
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        <div align=center> <b> No More Biofilm! </b> </div> <br>
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            <div align=center> <img src="http://2012.igem.org/wiki/images/9/92/Mr-clean.jpg" width="306" height="208"> </div>
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            <p>This year the CU iGEM team has harnessed the quorum sensing factors endogenous in the Salmonella enterica
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            serovar typhimurium LT2 strain to detect AHLs produced by other bacteria. Neither Salmonella nor E. coli have been
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            shown to produce detectable levels of AHLs, so they were model organisms for the detection of other AHL producing
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            bacteria such as ……. AHLs are small molecules synthesized by most gram negative bacteria, and are able to freely
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            diffuse throughout the membrane. When the concentration of AHL producing bacteria in a specific area increases, the
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            total concentration of AHLs diffusing into the cytoplasm also increases. Once a threshold level of AHLs are in the cytosol,
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            transcription factors like the Vibrio fisheri LuxR or Salmonella enterica SdiA activate gene synthesis for pathogenesis factors
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            to be produced. The SdiA transcription factor has been shown to be more sensitive to lower concentrations and a greater
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            diversity of AHLs than its LuxR homolog.</p>
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|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
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Latest revision as of 01:37, 4 October 2012

Boulder-landscape2012-team Boulder2012-project Boulder2012-results Boulder2012-model Boulder-panoramic2012
Boulder-landscape20121 Boulder-2012-project2 Boulder-2012-results3 Boulder-2012-model4 Boulder-2012-safety5 Boulder-2012-Attributions6

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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.



Quorum STOPPING



    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.
    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 Project page.

The video below is an animated overview of quorum sensing.