ses9_ln

ses9_ln - 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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Session 9 Lecture Notes 1. The authors are using a cell surface display method to screen a library of mutagenized OmpT – a bacterial cell surface protease- for new variants with either altered specificity or enhanced activity. 2. The library is formed using random mutagenesis via EP-PCR using Mn conc variation. They state that they don’t know how much mutation is going to be beneficial for the protein, so they chose this method. 3. FRET stands for Forster (or fluorescence) resonance energy transfer, which may occur between two chromophores in close proximity (<10nm). This is particularly useful in detecting the products of biochemical or enzymatic reactions. Can be done with small molecules or entire proteins (cyan fluorescent protein and Yellow fluorescent protein.) 4. FACS stands for fluorescent activated cell sorting and allows for cells to be
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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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ses9_ln - MIT OpenCourseWare http:/ocw.mit.edu 7.344...

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