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Staggered Extension Process (StEP) In Vitro Recombination

Aguinaldo, Anna Marie and Arnold, Frances H. (2003) Staggered Extension Process (StEP) In Vitro Recombination. In: Directed Evolution Library Creation: Methods and Protocols. Methods in Molecular Biology. No.231. Humana Press , Totowa, NJ, pp. 105-110. ISBN 159259395X.

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In vitro polymerase chain reaction (PCR)-based recombination methods are used to shuffle segments from various homologous DNA sequences to produce highly mosaic chimeric sequences. Genetic variations created in the laboratory or existing in nature can be recombined to generate libraries of molecules containing novel combinations of sequence information from any or all of the parent template sequences. Evolutionary protein design approaches, in which libraries created by in vitro recombination methods are coupled with screening (or selection) strategies, have successfully produced variant proteins with a wide array of modified properties including increased drug resistance (1,2), stability (3, 4, 5, 6), binding affinity (6), improved folding and solubility (7), altered or expanded substrate specificity (8,9), and new catalytic activity (10).

Item Type:Book Section
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Arnold, Frances H.0000-0002-4027-364X
Additional Information:© Humana Press Inc., Totowa, NJ 2003.
Subject Keywords:Negative Control Reaction; Increase Drug Resistance; StEP Recombination; Parental Template; DpnI Restriction Endonuclease
Series Name:Methods in Molecular Biology
Issue or Number:231
Record Number:CaltechAUTHORS:20180822-140726328
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:89039
Deposited By: George Porter
Deposited On:22 Aug 2018 22:58
Last Modified:03 Oct 2019 20:12

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