Tucker Ashley T, Powers Matthew J, Trent M Stephen, Davies Bryan W
Department of Molecular Biosciences, University of Texas at Austin, Austin, TX, USA.
Department of Infectious Diseases, University of Georgia, College of Veterinary Medicine, Athens, GA, USA.
Methods Mol Biol. 2019;1946:107-113. doi: 10.1007/978-1-4939-9118-1_11.
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.
鲍曼不动杆菌能迅速获得抗生素耐药性，其基因组编码耐受杀菌剂和干燥的机制，从而增强其在医院环境中的生存能力。需要能够快速剖析鲍曼不动杆菌基因组的工具，以便在遗传和机制层面了解促成其适应能力的细胞因子。虽然大量临床数据记录了鲍曼不动杆菌作为一种耐药病原体在全球范围内的增多，但剖析其分子细节的遗传工具却很有限。本方法描述了一种用于鲍曼不动杆菌靶向基因组编辑的重组介导基因工程（重组工程）系统。该系统可对多种基因和操纵子进行定向诱变，并在鲍曼不动杆菌的各种菌株中具有广泛应用。
