ses10_slides

ses10_slides - How does the enrichment factor compare to...

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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 using yeast surface display Boder, E.T.; Wittrup, K.D. Yeast surface display for screening combinatorial polypeptide libraries. Nat. Biotechnol. 1997 , 15 , 553- 557. Shiraga, S.; Kawakami, M.; Ishiguro, M.; Ueda, M. Enhanced reactivity of Rhizopus oryzae lipase displayed on yeast cell surfaces in organic solvents: potential as a whole-cell biocatalyst in organic solvents. App. Environ. Microbiol. 2005 , 71(8) , 4335-4338.
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Wittrup Lab yeast surface display • What are the limitations the authors identify with previous display methods that make their system attractive? What kinds of proteins are hard to express? • Explain the display scaffold. How are proteins displayed? (Figure 1) • How is the surface expression of scFvs verified? (Figure 2) How is the number of fusions quantified?
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Unformatted text preview: How does the enrichment factor compare to previously discussed methods? Explain their library generation and selection strategy. What are the results? What are the benefits/downsides to using this method? Figure 1- Aga-based protein display Image removed due to copyright restrictions. Please see Fig. 1 in Boder, E. T., and K. D. Wittrup. Yeast surface display for screening combinatorial polypeptide libraries. Nat. Biotechnol. 15(1997): 553-557. Figure 2 characterization of yeast surface fusions Image removed due to copyright restrictions. Please see Fig. 2 in Boder, E. T., and K. D. Wittrup. Yeast surface display for screening combinatorial polypeptide libraries. Nat. Biotechnol. 15(1997): 553-557. Rhizopus oryzae lipase display What are the authors trying to do and why is this significant? What is the strategy used? How is the enzyme library made and screened? What are the results? (Table 2) What are the pros/cons of the method?...
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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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ses10_slides - How does the enrichment factor compare to...

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