CHEN 3701 Lecture 30: 2015_11_2 Lecture
8 Nov 2015
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Goal: identify proteins(s) with new or improved function. Synthesize library of genes (random binding site sequence) One way to do this is called phage display put 1 gene into 1 cell protein is bound to cell in which we put gene advantage: efficient disadvantage: come from prokaryotic host --> limited quality. -> can switch to yeast display (eukaryotic host --> advantage) disadvantage: less efficient, cell size limits "library size) (10 7 - 109) ribosome/mrna: advantage: efficiency (10 10-13) disadvantage: acellular translation. Random diversity (a,c,t,g at all 3 codon sites) ex: for 8 runs, 68% Biased (elimate a from 3rd codon site): p = Generic code is conservative if make mutations, hydrophylic will likely stay hydrophylic and same for hydropohbic. Chen3701_biomolecularengineering page 1 advantageous to stabilize protein before. Simplistic model: mutations are beneficial, neutral, or eliminate activity (at least beneficial)*(remainder neutral); multiply that by # combinations. 2nd # = amount of beneficial mutations can also add in hindrance w/o elimination.
