关键词: directed evolution error prone PCR in vivo cloning lambda red proteins mutant libraries

Mesh : Bacillus / genetics Gene Library Mutagenesis Mutation Plasmids / genetics Polymerase Chain Reaction / methods Transformation, Genetic

来  源:   DOI:10.4161/bbug.1.5.12942   PDF(Sci-hub)

Abstract:
A simple and efficient method utilizing in vivo recombination to create recombinant libraries incorporating the products of PCR amplification is described. This will be especially useful for generating large pools of randomly mutagenized clones after error-prone PCR mutagenesis. Here we investigate various parameters to optimize this approach and we demonstrate that as little as 1 pmole of PCR fragment can generate a library with greater than 104 clones in a single transformation without ligation.
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