Generation of a Transposon Mutant Library in Staphylococcus aureus and Staphylococcus epidermidis Using bursa...

Transposon mutagenesis is a genetic process that involves the random insertion of transposons into a genome resulting in the disruption of function of the genes in which they insert. Identification of the insertion sites through DNA sequencing allows for the identification of the genes disrupted and the creation of "libraries" containing a collection of mutants in which a large number of the nonessential genes have been disrupted. These mutant libraries have been a great resource for investigators to understand the various biological functions of individual genes, including those involved in metabolism, antibiotic susceptibility, and pathogenesis. Here, we describe the detailed methodologies for constructing a sequence defined transposon mutant library in both Staphylococcus aureus and S. epidermidis using the mariner-based transposon, bursa aurealis.