Identification of Virulence Determinants During Host-Pathogen Interaction Using Tn-Seq Technology

Methods Mol Biol. 2020:2069:155-175. doi: 10.1007/978-1-4939-9849-4_12.

Abstract

Transposon sequencing (Tn-seq) is a powerful genetic tool that enables the detection of essential genes within a given environment. The application of Tn-seq to Staphylococcus aureus has generated transposon libraries in numerous strains with inactivation of virtually every nonessential gene in the genome. This exciting technology coupled with increasingly available computational tools has been deployed in animal models of infection to identify essential S. aureus genes within specific host environments. In this chapter, we describe the application of Tn-seq to a murine model of osteomyelitis as a paradigm for using this powerful technology to elucidate mechanisms of bacterial pathogenesis.

Keywords: In vivo; Invasive infection; Pathogenesis; S. aureus; Tn-seq osteomyelitis; Transposon sequencing.

MeSH terms

  • Animals
  • DNA Transposable Elements*
  • Gene Library*
  • High-Throughput Nucleotide Sequencing*
  • Mice
  • Staphylococcal Infections* / genetics
  • Staphylococcal Infections* / metabolism
  • Staphylococcus aureus* / genetics
  • Staphylococcus aureus* / metabolism
  • Staphylococcus aureus* / pathogenicity
  • Virulence Factors / genetics
  • Virulence Factors / metabolism

Substances

  • DNA Transposable Elements
  • Virulence Factors