Team:Calgary

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Revision as of 21:43, 15 July 2010

Welcome to the University of Calgary iGEM 2010 Wiki! As you can see, it's currently undergoing construction. Keep checking back as we continue to update!

We'll get something up very soon. Really. No joke. By very soon, we mean within the next month. Oh look! We kept our promise! Something's up! Enjoy!�

Preliminary Project Description

Many synthetic biology projects involve the expression of recombinant proteins in microorganisms such as E. coli. Although widely used, problems with expression are very common and it can be difficult to pinpoint where the problem lies. The goal of the University of Calgary 2010 iGEM team is to build a protein expression "troubleshooting kit". This kit will contain two plasmids into which non-functional genes can be cloned in. The subsequent cell growth will produce a fluorescent protein indicative of where protein expression is failing.

Protein expression happens in three steps: the transcription of the DNA to mRNA, the translation of mRNA into an amino acid sequence, and the folding of that amino acid sequence into a protein. Our system detects possible errors in these three steps using two circuits. The first circuit has a fluorescent reporter that is produced when DNA is transcribed into mRNA and another that is produced when mRNA is translated into a functional protein. When both reporter proteins are expressed in the cell, it indicates both transcription and translation are successful. The second circuit involves reporter systems that are activated as a result of protein misfolding. Two native stress-activated promoters from E. coli were constructed to fluorescent reporters corresponding to periplasmic and cytoplasmic protein misfolding. If the protein of interest misfolds in either area of the cell, one of the promoters will be activated and the corresponding fluorescence will be observed.