Protocols for Screening Antimicrobial Peptides That Influence Virulence Gene Expression in Staphylococcus aureus

Methods Mol Biol. 2017:1548:387-394. doi: 10.1007/978-1-4939-6737-7_28.

Abstract

Compounds that inhibit virulence gene expression in bacterial pathogens have received increasing interest as possible alternatives to the traditional antibiotic treatment of infections. For the human pathogen Staphylococcus aureus, we have developed two simple assays based on reporter gene fusions to central virulence genes that are easily applicable for screening various sources of natural and synthetic peptides for anti-virulence effects. The plate assay is qualitative but simultaneously assesses the effect of gradient concentrations of the investigated compound, whereas the liquid assay is quantitative and can be employed to address whether a compound is acting on the central quorum sensing regulatory system, agr, that controls a large number of virulence genes in S. aureus.

Keywords: Anti-virulence; Compound screening; Peptides; RNAIII; S. aureus; agr quorum sensing; hla; spa; β-Galactosidase/β-lactamase reporter genes.

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Antimicrobial Cationic Peptides / pharmacology*
  • Drug Discovery* / methods
  • Drug Evaluation, Preclinical
  • Gene Expression
  • Gene Expression Regulation, Bacterial / drug effects*
  • Genes, Reporter
  • Quorum Sensing / drug effects
  • Staphylococcus aureus / drug effects*
  • Staphylococcus aureus / genetics*
  • Staphylococcus aureus / pathogenicity
  • Virulence / genetics*
  • beta-Galactosidase / genetics
  • beta-Lactamases / genetics

Substances

  • Anti-Bacterial Agents
  • Antimicrobial Cationic Peptides
  • beta-Galactosidase
  • beta-Lactamases