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ses6_slides - MIT OpenCourseWare http/ocw.mit.edu 7.344...

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MIT OpenCourseWare http://ocw.mit.edu 7.344 Directed Evolution: Engineering Biocatalysts Spring 2008 For information about citing these materials or our Terms of Use, visit: http://ocw.mit.edu/terms .
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Enzyme evolution by chemical complementation Baker, K.; Bleczinski, C.; Lin, H.; Salazar-Jimenez, G.; Sengupta, D.; Krane, S.; Cornish, V.W. Chemical complementation: a reaction- independent genetic assay for enzyme catalysis. Proc. Natl. Acad. Sci. USA 2002 , 99 (26), 16537-16542. Azizi, B.; Chang, E.I.; Doyle, D.F. Chemical complementation: small- molecule-based genetic selection in yeast. Biochem. Biophys. Res. Commun. 2003 , 306 , 774-780. Lin, H.; Tao, H.; Cornish, V.W. Directed evolution of a glycosynthase via chemical complementation. J. Am. Chem. Soc. 2004 , 126 , 15051-15059.
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Chemical complementation as developed in the Cornish lab What are the authors trying to do? How do they go about engineering their system of complementation? What enzyme do they use to test their system? How does this work? (Figure 2, panel B) What are the results? (Figure 3, panels B and C) How do the authors confirm that the change in transcription is caused by enzyme turnover? Is this strategy general? What are the benefits and pitfalls?
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