Genetic Manipulation of Staphylococci

The ability to genetically manipulate bacteria is essential to understanding gene/protein function in these organisms. While basic cloning has become routine in molecular biology, many still view the ability to make directed mutations as a daunting or intimidating task. To aid the staphylococcal research community, the goal of this treatise is to describe the method of allelic exchange using temperature-sensitive plasmids that we have used to successfully produce a variety of mutations including single nucleotide changes in the Staphylococcus aureus chromosome. In addition, this chapter provides extensive summaries to aid in the construction of mutations, complementation plasmids, as well as transcription and translation reporters.