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MIT OpenCourseWare 7.344 Directed Evolution: Engineering Biocatalysts Spring 2008 For information about citing these materials or our Terms of Use, visit: .
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Session 4 Lecture Notes 1. How does chemical mutagenesis occur? Take a look at the chemdraw of the alkylation (explain term) of amine group-containing bases and how this causes AT to GC and GC to AT transitions (S N 2-type displacement). Treatment of DNA with chemicals can result in purposeful incorporation of mutations. What are some other ways to chemically mutate DNA other than alkylation? UV light – thymine dimerization, nitrous acid – deamination of C, A, and G, formic acid depurination of DNA by breaking N-glycosyl bonds of purine bases, hydrazine deconstruction of pyrimidine rings. 2. What are the authors using this method to study? Proteolysis of fibrin using fibrinolytic enzyme. Fibrin is a protein involved in the clotting of blood, so this class of enzymes could help thrombosis. The authors tried to improve its activity in vitro. 3. What are the results? The specific activity of an identified mutant (h-mBsFE) is almost 100% increased over wild-type. 80% of the clones showed at least a
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This note was uploaded on 11/11/2011 for the course BIO 7.344 taught by Professor Bobsauer during the Spring '08 term at MIT.

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ses4_ln - MIT OpenCourseWare http:/ 7.344...

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