A Method for Characterizing the Type III Secretion System's Contribution to Pathogenesis: Homologous Recombination to Generate Yersinia pestis Type III Secretion System Mutants

Methods Mol Biol. 2017:1531:155-164. doi: 10.1007/978-1-4939-6649-3_13.

Abstract

The type III (T3S) secretion system of many gram-negative bacteria is a surface-exposed protein secretion apparatus used to directly inject bacterial effector molecules into eukaryotic cells. These effector molecules contribute to bacterial pathogenesis in many ways, and have been shown to be crucial for infectivity. Here, we describe a protocol for using homologous recombination to generate T3S system mutants to assess the role of different T3S system proteins in bacterial pathogenesis.

Keywords: Homologous recombination; Pathogenesis; Secretion; Translocation; Type III secretion; Yersinia pestis.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Blotting, Western
  • Disease Models, Animal
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • HeLa Cells
  • Homologous Recombination
  • Humans
  • Mice
  • Mutation
  • Plague / microbiology
  • Protein Transport
  • Type III Secretion Systems / genetics*
  • Type III Secretion Systems / metabolism*
  • Yersinia pestis / genetics*
  • Yersinia pestis / metabolism*
  • Yersinia pestis / pathogenicity

Substances

  • Bacterial Proteins
  • Type III Secretion Systems