Team:Minnesota/Project/UV Absorption2

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<h1>Caffeine Production in Yeast</h1><br><br>
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<h1>UV Absorption by Skin Microbes</h1><br>
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The iGEM Minnesota 2012 Team is seeking to use the tools of synthetic biology to create genetically modified yeast with the ability to synthesize caffeine. Using genes that code for caffeine biosynthesis proteins from Coffea arabica, we hope to create and subsequently insert a plasmid into Saccharomyces cerevisiae that will confer the yeast with the ability to create caffeine in the lab. Immediate goals include the creation of a caffeine-production-inducing BioBrick that we'll submit to the registry, as well as a new empty yeast plasmid backbone that future iGEM teams can use for their projects.
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Between 2 and 3 million cases of nonmelanoma skin cancers arise each year, making skin cancer the most prevalent form of cancer worldwide. With skin cancer such a prominent problem worldwide, we are looking into alternative methods to protect against UV radiation. Photosynthetic organisms create an array of compounds, called mycosporine like amino acids, which naturally protect them against UV radiation. The cyanobacterium, Anabaena variabilis, contains a cluster of four genes capable of producing two UV protective compounds, mycosporine-glycine and shinorine.  Utilizing BioBrick™ vector techniques, we intend to clone these genes out of A. variabilis and into Escherichia coli for characterization.  After characterization and determination of the efficiency of the genes in E. coli, we will clone the shinorine gene cluster into Staphylococcus epidermidis. Successful introduction of this gene cluster and expression of UV-protective compounds could be useful as a one-time-application alternative to currently marketed sunscreens.  
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In coffee plants, the metabolic pathway for caffeine involves the use of AMP, GMP, and IMP to produce theobromine, which is the immediate precursor to caffeine. An important intermediate in the pathway is xanthosine, which already occurs naturally in S. cerevisiae. What our team seeks to do is introduce the genes into yeast that can transform the xanthosine into caffeine, via the xanothosine -> 7-methlyxanothine -> 7-methylxanthine -> theobromine -> caffeine pathway. The two genes we're incorporating into our plasmid are XMT1 and DXMT1. We plan to construct our plasmid de novo from several segments of DNA that we'll ligate using Gibson Assembly. Once construction of the plasmid is complete, we'll transform into yeast and screen for caffeine production.
 
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Latest revision as of 17:04, 30 September 2012

Team:Minnesota - Main Style Template Team:Minnesota

UV Absorption by Skin Microbes


      Between 2 and 3 million cases of nonmelanoma skin cancers arise each year, making skin cancer the most prevalent form of cancer worldwide. With skin cancer such a prominent problem worldwide, we are looking into alternative methods to protect against UV radiation. Photosynthetic organisms create an array of compounds, called mycosporine like amino acids, which naturally protect them against UV radiation. The cyanobacterium, Anabaena variabilis, contains a cluster of four genes capable of producing two UV protective compounds, mycosporine-glycine and shinorine. Utilizing BioBrick™ vector techniques, we intend to clone these genes out of A. variabilis and into Escherichia coli for characterization. After characterization and determination of the efficiency of the genes in E. coli, we will clone the shinorine gene cluster into Staphylococcus epidermidis. Successful introduction of this gene cluster and expression of UV-protective compounds could be useful as a one-time-application alternative to currently marketed sunscreens.

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