ses6_ln - MIT OpenCourseWare http/ 7.344...

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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 6 Lecture Notes 1. The authors are trying to create general strategy for the reporting of enzyme activity. They use small molecule receptors that are bound to a transcription factor and a DNA binding protein to activate transcription of reporter genes (LacZ). 2. The complementation system is engineered by making a strain of yeast that encodes LexA-DHFR and B42-GR fusion proteins under control of the GAL1 promoter. The LacZ reporter plasmid (spectinomycin) is transformed in as is the vector containing the enzyme (kanamycin). 3. The Cornish group uses the cephalosporinase enzyme to test their system. Cleavage of the B-lactam bond in the cephalosporin results in expulsion of the leaving group at the C3 position and breaks the bond between Mtx and Dex. The substrate (Mtx-Cephem-Dex) is added to the growth media in varying concentrations.
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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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ses6_ln - MIT OpenCourseWare http/ 7.344...

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