Download Protein Engineering by Dan E. Robertson and Joseph P. Noel (Eds.) PDF

By Dan E. Robertson and Joseph P. Noel (Eds.)

This MIE quantity covers tools for a large number of subject matters between that are computational equipment, laboratory tools, enzyme optimization, binding proteins/antibodies, and screening applied sciences.

Table of Contents
-Methodology
-Applications-Opzimization and Screening
-Applications-Directed Evolution of Enzymatic Function
-Applications-Evolution of Biosynthetic Pathways
-Devices, Antibodies and Vaccines

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By Dan E. Robertson and Joseph P. Noel (Eds.)

This MIE quantity covers tools for a large number of subject matters between that are computational equipment, laboratory tools, enzyme optimization, binding proteins/antibodies, and screening applied sciences.

Table of Contents
-Methodology
-Applications-Opzimization and Screening
-Applications-Directed Evolution of Enzymatic Function
-Applications-Evolution of Biosynthetic Pathways
-Devices, Antibodies and Vaccines

Show description

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2 METHODS IN ENZYMOLOGY, VOL. 388 Copyright 2004, Elsevier Inc. All rights reserved. 00 4 methodology [1] Fig. 1. Oligonucleotide primer design. we described a very non-stochastic approach, Gene Site Saturation MutagenesisTM (GSSMTM),* which systematically explores minimally all possible single amino acid substitutions along a protein sequence (Fig. 1). 18–21 This comprehensive technique introduces point mutations into every position within a target gene using degenerate primer sets containing 32 or 64 codons to generate a complete library of variants.

M. Fernandez, and J. M. Short, Biotechniques 5, 376 (1987). [1] gene site saturation mutagenesis 9 of random clones were determined, it was noticed that the annealing of primers with fewer mismatches from the template sequence was occasionally slightly favored. Primers with no mismatches with the template DNA were up to twice as likely to anneal and extend as primers that had three mismatches at the site of mutagenesis. Primers with one or two mismatches annealed and extended with intermediate efficiency.

L. Iverson, Protein Eng. 12, 349 (1999). 18 J. M. Short, Patent US 6,171,820 B1 (1999). 19 J. M. Short, Patent US 6,562,594 B1 (2003). 20 J. Braman, C. Papworth, and A. Greener, Methods Mol. Biol. 57, 31 (1996). 21 M. P. Weiner, G. L. Costa, W. Schoettlin, J. Cline, E. Mathur, and J. C. Bauer, Gene 151, 119 (1994). S. and/or other countries. 16 [1] gene site saturation mutagenesis 5 Fig. 2. Reaction mechanism. improved include, but are not limited to, stability, substrate specificity, pH performance, enantioselectivity, binding affinity, protein folding, gene expression, and productivity.

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