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Noncontiguous SCHEMA Protein Recombination

Smith, Matthew A. and Arnold, Frances H. (2014) Noncontiguous SCHEMA Protein Recombination. In: Directed Evolution Library Creation: Methods and Protocols. Methods in Molecular Biology. No.1179. Humana Press , New York, NY, pp. 345-352. ISBN 9781493910526.

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SCHEMA is a method of designing protein recombination libraries that contain a large fraction of functional proteins with a high degree of mutational diversity. In the previous chapter, we illustrated the method for designing libraries by swapping contiguous sequence elements. Here, we introduce the NCR (“noncontiguous recombination”) algorithm to identify optimal designs for swapping elements that are contiguous in the 3-D structure but not necessarily in the primary sequence. To exemplify the method, NCR is used to recombine three fungal cellobiohydrolases (CBH1s) to produce a library containing more than 500,000 novel chimeric sequences.

Item Type:Book Section
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Arnold, Frances H.0000-0002-4027-364X
Additional Information:© Springer Science+Business Media New York 2014. The authors acknowledge funding from the Institute for Collaborative Biotechnologies through grant W911NF-09-D-0001 from the US Army Research Office and the National Central University, Taiwan, through a Cooperative Agreement for Energy Research Collaboration. M.A.S. is supported by a Resnick Sustainability Institute fellowship.
Group:Resnick Sustainability Institute
Funding AgencyGrant Number
Army Research Office (ARO)W911NF-09-D-0001
National Central University (Taiwan)UNSPECIFIED
Resnick Sustainability InstituteUNSPECIFIED
Subject Keywords:Protein engineering; Homologous recombination; SCHEMA; Noncontiguous recombination; NCR; Chimeragenesis
Series Name:Methods in Molecular Biology
Issue or Number:1179
Record Number:CaltechAUTHORS:20180822-140726128
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:89037
Deposited By: George Porter
Deposited On:22 Aug 2018 22:11
Last Modified:16 Nov 2021 00:31

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