Miniaturized whole-cell bacterial bioreporter assay for identification of quorum sensing interfering compounds

J Microbiol Methods. 2018 Nov:154:40-45. doi: 10.1016/j.mimet.2018.10.005. Epub 2018 Oct 6.

Abstract

The continuing emergence and spread of antibiotic-resistant bacteria is worrisome and new strategies to curb bacterial infections are being sought. The interference of bacterial quorum sensing (QS) signaling has been suggested as a prospective antivirulence strategy. The AI-2 QS system is present in multiple bacterial species and has been shown to be correlated with pathogenicity. To facilitate the discovery of novel compounds interfering with AI-2 QS, we established a high-throughput setup of whole-cell bioreporter assay, which can be performed in either 96- or 384-well format. Agonistic or antagonistic activities of the test compounds against Escherichia coli LsrB-type AI-2 QS system are monitored by measuring the level of β-galactosidase expression. A control strain expressing β-galactosidase in quorum sensing-independent manner is included into the assay for false-positive detection.

Keywords: Assay miniaturization; Autoinducer-2; Bioreporter strain; High throughput screening; Quorum sensing inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaeal Proteins / genetics
  • Bacteria / drug effects
  • Bacteria / genetics
  • Bacteria / metabolism*
  • Biological Assay / methods*
  • Carrier Proteins / drug effects
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Drug Discovery / methods*
  • Drug Resistance, Bacterial
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / drug effects
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Pentanones / antagonists & inhibitors
  • Quorum Sensing*
  • Signal Transduction
  • beta-Galactosidase / metabolism

Substances

  • 4,5-dihydroxypentane-2,3-dione
  • Archaeal Proteins
  • Carrier Proteins
  • Escherichia coli Proteins
  • LsrB protein, E coli
  • Pentanones
  • beta-Galactosidase