Acinetobacter baumannii rapidly acquires antibiotic resistance, and its genome encodes mechanisms to tolerate biocides and desiccation, enhancing its persistence in hospital settings. Tools to rapidly dissect the A. baumannii genome are needed to understand cellular factors that contribute to its resiliency at a genetic and mechanistic level. While a substantial amount of clinical data has documented the global rise of A. baumannii as an antibiotic-resistant pathogen, genetic tools to dissect its molecular details have been limited. This procedure describes a recombination-mediated genetic engineering (recombineering) system for targeted genome editing of A. baumannii. This system can perform directed mutagenesis on wide-ranging genes and operons and has broad application in various strains of A. baumannii.
Keywords: Acinetobacter baumannii; Gene knockout; Genetic engineering; Mutagenesis; Recombinase; Recombineering.