BIO 3102 Lecture 18: Lec 18 - Directed Evolution for Protein Engineering
7 Apr 2017
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Express proteins usually in bac/yeast system and plug dna in. Each library member is grown separately in population. Express protein and extract it from bac then screen it for whether it has improved function we"re looking for. Make own mutations via looking at protein structure. Viral dna coding region comes out of bac and is connected to bacterial surface again. Dna that encodes protein f interest is fused t one of the coat protein genes of the phage. Dna encoding the protein f interest is displayed on the surface of the phage. Phage particles are then harvested and selected for the displayed proteins with novel functions. This is enzyme engineering to create enzyme to degrade unwanted substrates. Want enzyme to bind (novel function) and breaks it apart into non-hazardous chunks. P = product selecting for catalytic activity. If enz doesn"t degrade into product we want, then phage will bind product and find which enzymes could degrade this substrate.
