Chemical and biochemical strategies for the randomization of protein encoding DNA sequences: library construction methods for directed evolution.

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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Neylon C

doi

10.1093/nar/gkh315

keywords:

subject

Has Abstract

pub_date

2004-02-27 00:00:00

pages

1448-59

issue

4

eissn

0305-1048

issn

1362-4962

pii

32/4/1448

journal_volume

32

pub_type

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