Duplication-Insertion Recombineering: a fast and scar-free method for efficient transfer of multiple mutations in bacteria.

ABSTRACT: We have developed a new λ Red recombineering methodology for generating transient selection markers that can be used to transfer mutations between bacterial strains of both Escherichia coli and Salmonella enterica. The method is fast, simple and allows for the construction of strains with several mutations without any unwanted sequence changes (scar-free). The method uses λ Red recombineering to generate a marker-held tandem duplication, termed Duplication-Insertion (Dup-In). The Dup-Ins can easily be transferred between strains by generalized transduction and are subsequently rapidly lost by homologous recombination between the two copies of the duplicated sequence, leaving no scar sequence or antibiotic resistance cassette behind. We demonstrate the utility of the method by generating se

SUBMITTER: Nasvall J 

PROVIDER: S-EPMC5389514 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature