Acinetobacter baumannii is characterized as a nosocomial, gram-negative, multidrug-resistant bacterium, which has gained increasing attention due to its prevalence in hospital settings and high mortality rates upon infection. Currently, a number of different protocols have been developed in attempts to genetically alter A. baumannii, including multidrug-resistant strains. Although the bacterium has an unusual ability to uptake exogenous DNA in the natural environments, within the laboratory setting, gene manipulation to study virulence properties can be challenging. In this chapter we describe a general protocol for modification of specific genes using homologous recombination and a counterselectable marker.
Keywords: Acinetobacter; Allelic exchange; Counterselection; Gene deletion; Gene replacement; Homologous recombination; Multidrug resistance.