Part:BBa_K731000:Experience
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UNIQde415b57bd3d9945-partinfo-00000000-QINU UNIQde415b57bd3d9945-partinfo-00000001-QINU
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tkalkus |
The Stanford-Brown 2013 iGEM team further characterized and used this part as a positive control for an experimental ancestral version of CysE. To characterize it, we rescued a CysE knockout strain of E. coli grown on M9 minimal media plates. This gene is essential to the production of cysteine, an amino acid that does not occur abiotically. There is no cysteine in M9 minimal media, so any colonies growing on the plate must have the gene. We used a pUC19 vector with ampicillin resistance, and the knockout cells had kanamycin resistance. Because of the minimal media and dual antibiotics, any cells growing were almost guaranteed to have rescued the cell. There was sheet growth on our plates and we had the colonies sequenced to confirm the desired part was there. Indeed, the Trento 2012 CysE gene worked in rescuing our cells. Because the Trento team had taken growth curves to characterize their transformed cells, we also ran a day long growth curve test with the rescued cells in LB. |
