Creation of staphylococcal mutant libraries using transposon Tn917

Abstract

Non-directed mutagenesis of the staphylococcal genome is a global approach that can be used to identify the genetic basis for phenotypes of interest as well as for identifying regulators of gene expression. One such approach that has been widely used in the study of S. aureus and S. epidermidis is transposon Tn917 mutagenesis to generate random libraries of mutants. This chapter describes the use of plasmid pLTV1 (containing Tn917-lac) to generate Tn917 transposon mutants. Through the use of temperature manipulation and antibiotic selection, staphylococcal strains harboring this plasmid can be effectively mutagenized to create random libraries amenable to subsequent phenotypic screening and identification of transposon insertion sites.