Chemical and biochemical strategies for the randomization of protein encoding DNA sequences: library construction methods for directed evolution
Chemical and biochemical strategies for the randomization of protein encoding DNA sequences: library construction methods for directed evolution
Directed molecular evolution and combinatorial methodologies are playing an increasingly important role in the field of protein engineering. The general approach of generating a library of partially randomized genes, expressing the gene library to generate the proteins the library encodes and then screening the proteins for improved or modified characteristics has successfully been applied in the areas of protein-ligand binding, improving protein stability and modifying enzyme selectivity. A wide range of techniques are now available for generating gene libraries with different characteristics. This review will discuss these different methodologies, their accessibility and applicability to non-expert laboratories and the characteristics of the libraries they produce. The aim is to provide an up to date resource to allow groups interested in using directed evolution to identify the most appropriate methods for their purposes and to guide those moving on from initial experiments to more ambitious targets in the selection of library construction techniques. References are provided to original methodology papers and other recent examples from the primary literature that provide details of experimental methods.
1448-1459
Neylon, Cameron
697f067b-db25-4c41-9618-28f4b74f73aa
Neylon, Cameron
697f067b-db25-4c41-9618-28f4b74f73aa
Neylon, Cameron
(2003)
Chemical and biochemical strategies for the randomization of protein encoding DNA sequences: library construction methods for directed evolution.
Nucleic Acids Research, 32 (4), .
(doi:10.1093/nar/gkh315).
(Submitted)
Abstract
Directed molecular evolution and combinatorial methodologies are playing an increasingly important role in the field of protein engineering. The general approach of generating a library of partially randomized genes, expressing the gene library to generate the proteins the library encodes and then screening the proteins for improved or modified characteristics has successfully been applied in the areas of protein-ligand binding, improving protein stability and modifying enzyme selectivity. A wide range of techniques are now available for generating gene libraries with different characteristics. This review will discuss these different methodologies, their accessibility and applicability to non-expert laboratories and the characteristics of the libraries they produce. The aim is to provide an up to date resource to allow groups interested in using directed evolution to identify the most appropriate methods for their purposes and to guide those moving on from initial experiments to more ambitious targets in the selection of library construction techniques. References are provided to original methodology papers and other recent examples from the primary literature that provide details of experimental methods.
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Submitted date: 2003
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Local EPrints ID: 26767
URI: http://eprints.soton.ac.uk/id/eprint/26767
ISSN: 0305-1048
PURE UUID: b6c26fef-808f-444b-b714-015895e3232f
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Date deposited: 11 Apr 2006
Last modified: 15 Mar 2024 07:13
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Author:
Cameron Neylon
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