Difference between revisions of "Team:Austin UTexas"

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= UT Austin iGEM 2015 Home =
<h2> UT Austin iGEM 2015 Home </h2>
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<a href="http://utexas.edu/">University of Texas at Austin home</a>
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== BREAKING IS BAD ==
<h4>Project Description</h4>
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Our iGEM team, under the supervision of the Barrick lab at the University of Texas at Austin, developed five individual projects inspired by members’ interests and concerns in synthetic biology, with a foundation of technical skills and lab experience built during a spring semester course. The projects our team members have devised focus on a multitude of topics, from attempts at improving the stability and efficiency of existing genetic machines, to identifying bacterial factories that can have ecological function. Our projects focused on improving and expanding on the existing microbial factories in E. coli include an attempt to optimize the ΔguaB pDCAF strain of <i>E. coli</i> (Quandt <i>et al.</i>, 2013) to discount nutrients provided by non-caffeine methylxanthines, and a project assessing the evolutionary stability of yellow fluorescent protein, enhanced yellow fluorescent protein, and super-folder yellow fluorescent protein. Our projects that have a more ecological significance focus on drought, and the introduction/amplification of genes that produce trehalose, auxins, and ACC deaminase; transformation of the bee gut bacteria <i>Snodgrassella</i> and <i>Gilliamella</i> with the NHase gene to degrade the neonicotinoid thiacloprid; and the use of selective enrichment to isolate a strain/strains of bacteria that can degrade the neonicotinoid thiamethoxam. An additional project with more direct human impact concerns itself with creating a food-safe bacterial pH sensor to detect when milk has spoiled.</br>
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<b>References</b></br>
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<p><h5>Quandt, Erik M., et al. "Decaffeination and measurement of caffeine content by addicted Escherichia coli with a refactored N-demethylation operon from Pseudomonas putida CBB5." ACS synthetic biology 2.6 (2013): 301-307.</br></h5>
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<p><h5>Zhang, Hui-Juan, et al. "Biotransformation of the neonicotinoid insecticide thiacloprid by the bacterium Variovorax boronicumulans strain J1 and mediation of the major metabolic pathway by nitrile hydratase."Journal of agricultural and food chemistry 60.1 (2011): 153-159.</br></h5>
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<p><h5>Liu, Juan, et al. "An improved method for extracting bacteria from soil for high molecular weight DNA recovery and BAC library construction." The Journal of Microbiology 48.6 (2010): 728-733.</br></h5>
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[[Image:2015_Austin_UTexas_homepage-BB.png|300px| right]]
  
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After an organism is reprogrammed with a genetic device, the device will often mutate, or “break”, decreasing the metabolic load on the organism and giving it a competitive advantage. This commonly allows the organism with the broken genetic device to dominate the population, undermining the purpose of the original reprogramming. We measured how quickly several plasmids encoding different fluorescent proteins broke during laboratory culturing and endeavored to identify and better characterize the types of sequences that are prone to breaking. We found that certain devices broke more quickly and characterized the mutations that caused loss of function. We then expanded on this research by transforming four ''E. coli'' strains with fluorescent protein plasmids. Breaking times varied noticeably between strains, suggesting that the host’s own genetic material also influenced device stability. Finally, we took part in the interlab measurement study and found that one of these plasmids was also very unstable.
  
<h4> Editing your wiki </h4>
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: '''[[Team:Austin_UTexas/Project/Problem | PROBLEM: GENETIC INSTABILITY]]'''
<p>On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world! </p>
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<p> <a href="https://2015.igem.org/wiki/index.php?title=Team:Austin_UTexas&action=edit"> Click here to edit this page! </a></p>
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<p>See tips on how to edit your wiki on the <a href="https://2015.igem.org/TemplatesforTeams_Code_Documentation">Template Documentation</a> page.</p>
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: '''[[Team:Austin_UTexas/Project/Plasmid_Study | PART 1: OBSERVING FLUORESCENT GENE STABILITY]]'''
  
<h4>Templates </h4>
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: '''[[Team:Austin_UTexas/Project/Strain_Study | PART 2: GENETIC STABILITY IN DIFFERENT STRAINS]]'''
<p> This year we have created templates for teams to use freely. More information on how to use and edit the templates can be found on the
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<a href="https://2015.igem.org/TemplatesforTeams_Code_Documentation">Template Documentation </a> page.</p>
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: '''[[Team:Austin_UTexas/Interlab_Study | PART 3: BREAKING IS BAD FOR THE INTERLAB STUDY]]'''
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== CAFFEINATED COLI II ==
  
<h4> Uploading pictures and files </h4>
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[[Image:2015_Austin_UTexas_homepage-caffeine.png|250px|right]]
<p> You can upload your pictures and files to the iGEM 2015 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. <br />
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When you upload, set the "Destination Filename" to <code>Team:YourOfficialTeamName/NameOfFile.jpg</code>. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)</p>
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<a href="https://2015.igem.org/Special:Upload">CLICK HERE TO UPLOAD FILES</a>
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The 2012 UT Austin iGEM team developed the Δ''guaB'' pDCAF3 strain that could measure the concentration of caffeine by degrading it into a viable replacement for guanine. We extended this work in two ways. First, we redesigned the pDCAF3 plasmid for greater genetic stability and to make it BioBrick compatible. Second, we created a collection of plasmids containing all subsets of the component enzymes to enable different methylxanthines to be degraded with high specificity. These plasmids could potentially be used to more accurately measure the amounts of different methylxanthines in a beverage. For example, coffee contains mainly caffeine, but tea and cocoa contains mixtures of caffeine, theobromine, and theophylline.
  
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: '''[[Team:Austin_UTexas/Project/Caffeine | PART 4: REDESIGNING DECAFFEINATION PLASMIDS]]'''
  
 
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| rowspan="2" |  [[Image:UT_Austin_CSSB.png|link=http://cssb.utexas.edu|220px]]
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| rowspan="2" |  [[Image:UT_Austin_BEACON.png|150px|link=http://beacon-center.org]]
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| [[Image:UT_Austin_ICMB.png|link=http://icmb.utexas.edu|300px]]
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| rowspan="2" | [[Image:UT_Austin_FRI.png|link=https://cns.utexas.edu/fri|200px]]
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| [[Image:UT_Austin_MBS.png|link=http://molecularbiosci.utexas.edu|300px]]
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{{Austin_UTexas_Footer}}

Latest revision as of 02:12, 19 September 2015

UT Austin iGEM 2015 Home

BREAKING IS BAD

2015 Austin UTexas homepage-BB.png

After an organism is reprogrammed with a genetic device, the device will often mutate, or “break”, decreasing the metabolic load on the organism and giving it a competitive advantage. This commonly allows the organism with the broken genetic device to dominate the population, undermining the purpose of the original reprogramming. We measured how quickly several plasmids encoding different fluorescent proteins broke during laboratory culturing and endeavored to identify and better characterize the types of sequences that are prone to breaking. We found that certain devices broke more quickly and characterized the mutations that caused loss of function. We then expanded on this research by transforming four E. coli strains with fluorescent protein plasmids. Breaking times varied noticeably between strains, suggesting that the host’s own genetic material also influenced device stability. Finally, we took part in the interlab measurement study and found that one of these plasmids was also very unstable.

PROBLEM: GENETIC INSTABILITY
PART 1: OBSERVING FLUORESCENT GENE STABILITY
PART 2: GENETIC STABILITY IN DIFFERENT STRAINS
PART 3: BREAKING IS BAD FOR THE INTERLAB STUDY


CAFFEINATED COLI II

2015 Austin UTexas homepage-caffeine.png

The 2012 UT Austin iGEM team developed the ΔguaB pDCAF3 strain that could measure the concentration of caffeine by degrading it into a viable replacement for guanine. We extended this work in two ways. First, we redesigned the pDCAF3 plasmid for greater genetic stability and to make it BioBrick compatible. Second, we created a collection of plasmids containing all subsets of the component enzymes to enable different methylxanthines to be degraded with high specificity. These plasmids could potentially be used to more accurately measure the amounts of different methylxanthines in a beverage. For example, coffee contains mainly caffeine, but tea and cocoa contains mixtures of caffeine, theobromine, and theophylline.

PART 4: REDESIGNING DECAFFEINATION PLASMIDS



UT Austin CSSB.png UT Austin BEACON.png UT Austin ICMB.png UT Austin FRI.png
UT Austin MBS.png