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Project GluE coli

Taking advantage of muscle adhesive proteins (MAPs) really stuck on to this year’s UT iGEM team. Muscle adhesive proteins have amassed much attention as a potential adhesive for biomedical, underwater and other commercially relevant applications. Furthermore, muscle adhesive proteins are sought for their biodegradability, biocompatibility and ability to adhere to various substrates. However, production of muscle adhesive proteins proves to be an arduous task. Extraction-based production and in-vitro-based production are expensive, inefficient, and unsustainable. Thus, our team focused on improving the efficiency of in vivo production of MAPs utilizing fusion protein fp-151. MAPs derive their adhesive properties from the hydroxylation of tyrosine to L-3,4-dihydroxyphenylalanine (L-DOPA) then to dopaquinone. Our team employed an amber suppression system in order to incorporate an unnatural amino acid, in this case L-DOPA, at the stop codon sequences. Our team aims to develop the technology for large-scale MAP production in E. coli that will ultimately allow for rapid, cost-effective, and commercially viable production of an adhesive for biomedical applications.

Project ZombiE.coli

UT’s ZombiE.coli project aims to a develop a tightly regulated genetic switch that is triggered by bacterial quorum signaling and leads to feed-forward propagation of the genetic output in the form of red or green fluorescence as well as amplification of quorum signaling. The switch relies on simple one-way Cre/loxP recombination combined with native quorum signaling to provide us with a system that models transmissible disease spread between populations. We have likened this to an airborne zombie epidemic, in which an “infected” zombie cell is capable of restructuring the genes of a normal cell, turning it into a flesh-hungry counterpart. This system will be useful not only as a simple disease outbreak model for intermediate-level biology education, but also, could provide new insights to how bacterial populations communicate in three dimensions and under different genetic backgrounds.

Project PopeyE.coli

In an effort to improve the efficiency, ease, and quality of promoter and RBS strength measurements, we focused on developing a dual fluorescence reporter for simultaneous monitoring both transcription and translation. To measure both processes separately, two fluorescent reporters, the Spinach aptamer and mCherry red fluorescent protein, were assembled into a single construct. The Spinach-mCherry dual reporter is a unique concept; Spinach is a short RNA aptamer that binds to its ligand, DFHBI, and allows it to emit green fluorescence similar to GFP. This gives insight into the direct production of the mCherry-encoding mRNA without the need to wait for protein folding and maturation of the fluorophore. This technique attempted to expand upon current efforts to measure promoter strength relative to a reference standard used by the iGEM community.