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Random-priming in vitro recombination: an effective tool for directed evolution

Shao, Zhixin and Zhao, Huimin and Giver, Lori and Arnold, Frances H. (1998) Random-priming in vitro recombination: an effective tool for directed evolution. Nucleic Acids Research, 26 (2). pp. 681-683. ISSN 0305-1048. PMCID https://www.ncbi.nlm.nih.gov/pmc/articles/PMC147287/. http://resolver.caltech.edu/CaltechAUTHORS:SHAnar98

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Abstract

A simple and efficient method for in vitro mutagenesis and recombination of polynucleotide sequences is reported. The method involves priming template polynucleotide(s) with random-sequence primers and extending to generate a pool of short DNA fragments which contain a controllable level of point mutations. The fragments are reassembled during cycles of denaturation, annealing and further enzyme-catalyzed DNA polymerization to produce a library of full-length sequences. Screening or selecting the expressed gene products leads to new variants with improved functions, as demonstrated by the recombination of genes encoding different thermostable subtilisins in order to obtain enzymes more stable than either parent.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://www.ncbi.nlm.nih.gov/pmc/articles/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC147287//PubMed CentralArticle
ORCID:
AuthorORCID
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 1998 by Oxford University Received September 15, 1997; Revised and Accepted November 21, 1997 This research is supported by Eli Lilly and Co., the US Office of Naval Research (N0014-96-1-340 and N00014-97-1-0433), the US Department of Energy’s program in Biological and Chemical Technologies Research within the Office of Industrial Technologies, Energy Efficiency and Renewables. Z.S. is grateful to Drs Peggy Arps and Kentaro Miyazaki for helpful discussions and to Michaeleen Callahan for excellent technical assistance.
Funders:
Funding AgencyGrant Number
Eli LillyUNSPECIFIED
Office of Naval Research (ONR)N0014-96-1-340
Office of Naval Research (ONR)N00014-97-1-0433
Department of Energy (DOE)UNSPECIFIED
PubMed Central ID:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC147287/
Record Number:CaltechAUTHORS:SHAnar98
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:SHAnar98
Alternative URL:http://dx.doi.org/10.1093/nar/26.2.681
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:815
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:08 Oct 2005
Last Modified:13 Sep 2017 16:16

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