Batch transduction of transposon mutant libraries for rapid phenotype screening in staphylococcus

Abstract

In the gram-positive pathogen Staphylococcus aureus, transposon mutagenesis is a useful method of screening large numbers of mutants for a given phenotype. However, constructing a transposon mutant library can take several months of work and validation in the laboratory. In this chapter, we describe a method for batch transduction of existing transposon mutations into new genetic backgrounds. Transduction in S. aureus is accomplished quickly and easily in most commonly used laboratory strains. The method described herein utilizes transduction to facilitate the rapid creation of new libraries and quick screening in strains containing phenotypic reporter constructs.