Genetic manipulation of Staphylococcus aureus

Curr Protoc Microbiol. 2014 Feb 6:32:Unit 9C.3.. doi: 10.1002/9780471729259.mc09c03s32.

Abstract

Staphylococcus aureus is a facultative anaerobic Gram-positive coccus and a member of the normal skin flora, as well as that of the nasal passages of humans. However, S. aureus can also gain entry into the host and cause life-threatening infections or persist as disease foci that develop into suppurative abscesses. While genetically tractable, the manipulation of S. aureus remains challenging. This unit describes methods developed in our laboratory for gene disruption by allelic replacement and transposition. We also provide a protocol for bacteriophage-mediated transduction of mutants marked with selectable alleles and describe plasmid utilization for complementation studies.

Keywords: Staphylococcus aureus; allelic replacement; bursa aurealis; plasmid; transduction.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA Transposable Elements
  • Humans
  • Mutagenesis, Insertional / methods*
  • Plasmids / genetics
  • Polymerase Chain Reaction / methods*
  • Staphylococcal Infections / microbiology*
  • Staphylococcus aureus / classification
  • Staphylococcus aureus / genetics*
  • Staphylococcus aureus / isolation & purification
  • Transduction, Genetic / methods*

Substances

  • DNA Transposable Elements