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Combinatorial Recombination of Gene Fragments to Construct a Library of Chimeras

Farrow, Mary F. and Arnold, Frances H. (2010) Combinatorial Recombination of Gene Fragments to Construct a Library of Chimeras. Current Protocols in Protein Science, 62 (1). 26.2.1-26.2.20. ISSN 1934-3655. http://resolver.caltech.edu/CaltechAUTHORS:20180820-160726572

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Abstract

Recombination of distantly related and nonrelated genes is difficult using traditional PCR‐based techniques, and truncation‐based methods result in a large proportion of nonviable sequences due to frame shifts, deletions, and insertions. This unit describes a method for creating libraries of chimeras through combinatorial assembly of gene fragments. It allows the experimenter to recombine genes of any identity and to select the sites where recombination takes place. Combinatorial recombination is achieved by generating gene fragments with specific overhangs, or sticky ends. The overhangs permit the fragments to be ligated in the correct order while allowing independent assortment of blocks with identical overhangs. Genes of any identity can be recombined so long as they share 3 to 5 base pairs of identity at the desired recombination sites. Simple adaptations of the method allow incorporation of specific gene fragments.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/0471140864.ps2602s61DOIArticle
ORCID:
AuthorORCID
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2010 John Wiley & Sons, Inc. Issue Online: 01 August 2010; Version of Record online: 01 August 2010.
Subject Keywords:chimera; recombination; directed evolution; protein design; library creation; combinatorial library
Record Number:CaltechAUTHORS:20180820-160726572
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20180820-160726572
Official Citation:Farrow, M. F. and Arnold, F. H. (2010), Combinatorial Recombination of Gene Fragments to Construct a Library of Chimeras. Current Protocols in Protein Science, 62: 26.2.1-26.2.20. doi:10.1002/0471140864.ps2602s61
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:88958
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:20 Aug 2018 23:37
Last Modified:20 Aug 2018 23:37

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