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Project Description

This year’s iGEM competition season is an exciting one - one that will certainly unravel fresh insight into the realm of synthetic biology. With this in mind, the 2016 Lambert iGEM Team hopes to gain more insight into the degradation of chromoproteins and consequently characterize two well known degradation mechanisms - ClpXP and ClpAP.

Protein degradation plays a crucial role in maintaining homeostasis throughout the cell. When a protein is sentenced for destruction, it is tagged or somehow otherwise edited to notify proteases within the cell to degrade it. Perhaps the most well known and well studied protein degradation pathway is the SsrA RNA tagging system, in which ClpXP and ClpAP, two protease mechanisms, are reported to be an integral part of the degradation pathway. While these two proteases differ slightly in name, scientists are skeptical as to how they differ in function. According to studies conducted by numerous synthetic labs, chiefly the one at MIT, both ClpXP and ClpAP contribute to the degradation of proteins that contain an eleven peptide residue on the C-terminus (Gottesman, et al.). Moreover, both ClpXP and ClpAP identify SsrA-tagged proteins and denature them into a linear peptide strands which are later fed into ClpP, a common proteolytic subunit shared by both ClpXP and ClpAP. Because both proteases degrade proteins in such an indistinguishable manner, there has been widespread controversy among the synthetic biology community over which protease, ClpXP or ClpAP, contributes more to the degradation of proteins. While some studies state that the two proteases may vary in their affinities with different substrates, others show that ClpXP degrades SsrA tagged GFP more efficiently than ClpAP during the exponential phase of E. coli growth (Farrell, et al.).

Consequently, the Lambert iGEM Team has chosen to characterize ClpXP and ClpAP with regards to how both mechanisms degrade chromoproteins. More specifically, this year’s team will measure the time and subsequently the rate at which chromoproteins are degraded by both ClpXP and ClpAP. This will be achieved by designing a contraption in which a camera will capture footage of the light emitted by chromoproteins over a period of time; this data will later be quantified by using advanced photographic software to determine the time it took for the chromoprotein to lose its color (and consequently be degraded). In order to execute this project successfully, members of the Lambert iGEM Team have been preparing themselves both academically and physically. For example, thus far, the Lambert iGEM Team has been practicing basic laboratory skills (such as transformations, mini-preps, mock 3A assemblies) to prepare for building the actual construct. Moreover, the team has also been coordinating with Georgia Tech professor Mark Styczynski and his fellow graduate students to develop a comprehensive and competitive project. All in all, the 2016 Lambert iGEM Team is in full gear to prepare for the upcoming iGEM Jamboree.

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We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.

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