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• W1979921600 abstract "Focused mutant library generation methods have been developed to improve mainly “localizable” enzyme properties such as activity and selectivity. Current multi-site saturation methods are restricted by the gene sequence, require subsequent PCR steps and/or additional enzymatic modifications. Here we report, a multiple site saturation mutagenesis method, OmniChange, which simultaneously and efficiently saturates five independent codons. As proof of principle, five chemically cleaved DNA fragments, each carrying one NNK-degenerated codon, were generated and assembled to full gene length in a one-pot-reaction without additional PCR-amplification or use of restriction enzymes or ligases. Sequencing revealed the presence of up to 27 different codons at individual positions, corresponding to 84.4% of the theoretical diversity offered by NNK-degeneration. OmniChange is absolutely sequence independent, does not require a minimal distance between mutated codons and can be accomplished within a day." @default.
• W1979921600 created "2016-06-24" @default.
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• W1979921600 date "2011-10-19" @default.
• W1979921600 modified "2023-10-18" @default.
• W1979921600 title "OmniChange: The Sequence Independent Method for Simultaneous Site-Saturation of Five Codons" @default.
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• W1979921600 doi "https://doi.org/10.1371/journal.pone.0026222" @default.
• W1979921600 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3198389" @default.
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