Darwin Assembly: fast, efficient, multi-site bespoke mutagenesis.

ABSTRACT: Engineering proteins for designer functions and biotechnological applications almost invariably requires (or at least benefits from) multiple mutations to non-contiguous residues. Several methods for multiple site-directed mutagenesis exist, but there remains a need for fast and simple methods to efficiently introduce such mutations - particularly for generating large, high quality libraries for directed evolution. Here, we present Darwin Assembly, which can deliver high quality libraries of >108 transformants, targeting multiple (>10) distal sites with minimal wild-type contamination (<0.25% of total population) and which takes a single working day from purified plasmid to library transformation. We demonstrate its efficacy with whole gene codon reassignment of chloramphenicol acetyl tran

SUBMITTER: Cozens C 

PROVIDER: S-EPMC5934624 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature