Team:MSU-Michigan/Parts
Parts
The parts were taken from the Shewanella oneidensis MR-1 genome by finding a regulator on RegPrecise. Then the consensus sequence was located at the appropriate distance from a gene and DNA upstream from the gene was taken, which included the consensus sequence. This ensured that the ribosomal binding site would be in the sequence we took as well as any other necessary machinery for the following gene to be regulated by the promoter that we extracted. All of the promoters were then amplified using PCR and inserted into the prL814 plasmid in place of the T7A1 promoter.
Each part that the MSU iGEM team constructed, was taken from the Shewanella oneidensis MR-1 genome by finding a regulator for each desired part on http://regprecise.lbl.gov/RegPrecise/. These parts were all built to detect contaminants. A provided consensus sequence was then taken at the appropriate distance from a gene and included with the DNA that is upstream from that gene. This selection process ensured that the ribosomal binding site would be in the sequence taken, as well as any other necessary machinery for the following gene to be regulated by the promoter that was extracted. All of the promoters were then amplified using PCR and inserted into the prL814 plasmid in place of the T7A1 promoter. From the nine parts created ( part numbers listed below) Paraquat was the main part that the MSU team chose to pursue for GFP experiments, and proof of concept, due to no metabolic relevance to the cell.
Part Table
"Confirmation of promoters induced by (from left to right) Blue light, Paraquat, Nitrate (NO3), and Copper"
<groupparts>iGEM17 MSU-Michigan</groupparts>
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