Mutation of GTFs

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To induce favorable mutations we plan to employ directed mutagenesis on the aforementioned global transcription factors to increase current output. The mutations would be introduced by error prone PCR, which is a technique used to generate randomized genomic libraries. Error-prone PCR will allow us to initiate DNA amplification starting with tiny amounts of parent molecule to produce considerable amounts of the mutated gene. This technique is based on the principle that Taq polymerase is capable of annealing incompatible base-pairs to each other during amplification under imperfect PCR conditions. The figure below contrasts the external conditions between PCR and error prone PCR.

Error prone.jpg