Difference between revisions of "Team:Austin UTexas"

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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 culture 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 | PROBLEM: GENETIC INSTABILITY]]'''
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: '''[[Team:Austin_UTexas/Project/Problem | PROBLEM: GENETIC INSTABILITY]]'''
  
'''[[Team:Austin_UTexas/Project/Plasmid_Study | PART 1: MEASURING FLUORESCENT PROTEIN STABILITY]]'''
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: '''[[Team:Austin_UTexas/Project/Plasmid_Study | PART 1: BREAKING FLUORESCENT PROTEIN EXPRESSION]]'''
  
'''[[Team:Austin_UTexas/Project/Strain_Study | PART 2: STUDYING STABILITY IN DIFFERENT STRAINS]]'''
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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 ===
 
=== CAFFEINATED COLI ===
  
[[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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We constructed plasmids that are capable of degrading different methylxanthines and allowing us to accurately measure their concentrations.
  
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.
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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 the subsets of the component enzymes that enable degrading different methylxanthines with fairly high specificity. These plasmids could potentially be used to measure the content in a beverage of theobromine (at high concentrations in tea) and caffeine (at high concentrations in coffee), for example.
  
'''[[Team:Austin_UTexas/Project/Caffeine | PART 3: REDESIGNING DECAFFEINATION PLASMIDS]]'''
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: '''[[Team:Austin_UTexas/Project/Caffeine | PART 4: REDESIGNING DECAFFEINATION PLASMIDS]]'''
  
 
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Revision as of 19:58, 18 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 culture 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: BREAKING FLUORESCENT PROTEIN EXPRESSION
PART 2: GENETIC STABILITY IN DIFFERENT STRAINS
PART 3: BREAKING IS BAD FOR THE INTERLAB STUDY


CAFFEINATED COLI

2015 Austin UTexas homepage-caffeine.png

We constructed plasmids that are capable of degrading different methylxanthines and allowing us to accurately measure their concentrations.

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 the subsets of the component enzymes that enable degrading different methylxanthines with fairly high specificity. These plasmids could potentially be used to measure the content in a beverage of theobromine (at high concentrations in tea) and caffeine (at high concentrations in coffee), for example.

PART 4: REDESIGNING DECAFFEINATION PLASMIDS



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