A method for increasing electroporation competence of Gram-negative clinical isolates by polymyxin B nonapeptide.

ABSTRACT: The study of clinically relevant bacterial pathogens relies on molecular and genetic approaches. However, the generally low transformation frequency among natural isolates poses technical hurdles to widely applying common methods in molecular biology, including transformation of large constructs, chromosomal genetic manipulation, and dense mutant library construction. Here we demonstrate that culturing clinical isolates in the presence of polymyxin B nonapeptide (PMBN) improves their transformation frequency via electroporation by up to 100-fold in a dose-dependent and reversible manner. The effect was observed for PMBN-binding uropathogenic Escherichia coli (UPEC) and Salmonella enterica strains but not naturally polymyxin resistant Proteus mirabilis. Using our PMBN electroporation method

SUBMITTER: Qin J 

PROVIDER: S-EPMC9270391 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature