Degenerate Oligonucleotide Gene Shuffling

Show simple item record Bergquist, Peter en Gibbs, MD en
dc.contributor.editor Arndt, KM en
dc.contributor.editor Muller, KM en 2011-07-24T21:50:39Z en 2007 en
dc.identifier.citation In K. Mueller & K. Arndt (eds), Protein Engineering Protocols, Methods in Molecular Biology series, Totowa NJ: Humana Press. 2007 en
dc.identifier.isbn 1588290727 en
dc.identifier.isbn 9781588290724 en
dc.identifier.uri en
dc.description.abstract Improvement of the biochemical characteristics of enzymes has been aided by misincorporation mutagenesis and DNA shuffling. Shuffling techniques can be used on a collection of mutants of the same gene, or related families of genes can be shuffled to produce mutants encoding chimeric gene products. One difficulty with current shuffling procedures is the predominance of unshuffled (“parental”) molecules in the pool of mutants. We describe a procedure for gene shuffling using degenerate primers that allows control of the relative levels of recombination between the genes that are shuffled and reduces the regeneration of unshuffled parental genes. This procedure has the advantage of avoiding the use of endonucleases for gene fragmentation before shuffling and allows the use of random mutagenesis of selected segments of the gene as part of the procedure. We illustrate the use of the technique with a diverse family of β-xylanase genes that possess widely different G and C contents. en
dc.publisher Humana Press. en
dc.relation.ispartof Protein Engineering Protocols en
dc.relation.ispartofseries Methods in Molecular Biology series en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. en
dc.rights.uri en
dc.title Degenerate Oligonucleotide Gene Shuffling en
dc.type Book Item en
dc.identifier.doi 10.1385/1-59745-187-8:191 en
pubs.volume 352 en
dc.rights.holder Copyright: Humana Press Inc., Totowa, NJ en Totowa, NJ en
dc.rights.accessrights en
pubs.elements-id 11615 en
pubs.record-created-at-source-date 2010-09-01 en

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