Difference between revisions of "Team:Austin UTexas"

 
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= UT Austin iGEM 2015 Home =
== UT Austin iGEM 2015 Home ==
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=== BREAKING IS BAD ===
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== BREAKING IS BAD ==
  
 
[[Image:2015_Austin_UTexas_homepage-BB.png|300px| right]]  
 
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The 2015 UT Austin iGEM Team's major project is primarily concerned with the stability of genetically modified devices in bacteria. Devices run the risk of breaking, or mutating, if they prove to be too much metabolic stress to an organism. We endeavored to identify and better characterize the types of sequences that are prone to breaking, and used fluorescent-protein coding strains of <i>E. coli</i> to observe breaking speed.
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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.
  
[[Team:Austin_UTexas/Project/Problem | GO TO PROJECT PAGE]]
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: '''[[Team:Austin_UTexas/Project/Problem | PROBLEM: GENETIC INSTABILITY]]'''
  
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: '''[[Team:Austin_UTexas/Project/Plasmid_Study | PART 1: OBSERVING FLUORESCENT GENE STABILITY]]'''
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: '''[[Team:Austin_UTexas/Project/Strain_Study | PART 2: GENETIC STABILITY IN DIFFERENT STRAINS]]'''
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: '''[[Team:Austin_UTexas/Interlab_Study | PART 3: BREAKING IS BAD FOR THE INTERLAB STUDY]]'''
 
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=== CAFFEINATED COLI ===
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== CAFFEINATED COLI II ==
  
[[Image:2015_Austin_UTexas_homepage-caffeine.png|250px| right]]
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[[Image:2015_Austin_UTexas_homepage-caffeine.png|250px|right]]
  
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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.
  
We constructed plasmids that are capable of degrading different methylxanthines and allowing us to accurately measure their concentrations. The 2012 UT Austin iGEM team, with the help of Dr. Erik Quandt, developed the ΔguaB pDCAF3 strain that could measure the concentration of caffeine by degrading it into a viable replacement for guanine. The 2015 team modified the pDCAF3 strain into seven plasmids that could degrade different methylxanthines with fairly high specificity.<br>
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: '''[[Team:Austin_UTexas/Project/Caffeine | PART 4: REDESIGNING DECAFFEINATION PLASMIDS]]'''
<a href="https://2015.igem.org/Team:Austin_UTexas/Project/Caffeine"><font color="017e70"><b>GO TO CAFFEINE PAGE</b></font></a>
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<b>References</b></br>
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<h6>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></h6>
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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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| rowspan="2" | [[Image:UT_Austin_FRI.png|link=https://cns.utexas.edu/fri|200px]]
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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