Intra-species bacterial quorum sensing studied at single cell level in a double droplet trapping system

Int J Mol Sci. 2013 May 21;14(5):10570-81. doi: 10.3390/ijms140510570.

Abstract

In this paper, we investigated the intra-species bacterial quorum sensing at the single cell level using a double droplet trapping system. Escherichia coli transformed to express the quorum sensing receptor protein, LasR, were encapsulated in microdroplets that were positioned adjacent to microdroplets containing the autoinducer, N-(3-oxododecanoyl)-L-homoserine lactone (OdDHL). Functional activation of the LasR protein by diffusion of the OdDHL across the droplet interface was measured by monitoring the expression of green fluorescent protein (GFP) from a LasR-dependent promoter. A threshold concentration of OdDHL was found to induce production of quorum-sensing associated GFP by E. coli. Additionally, we demonstrated that LasR-dependent activation of GFP expression was also initiated when the adjacent droplets contained single E. coli transformed with the OdDHL synthase gene, LasI, representing a simple quorum sensing circuit between two droplets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Butyrolactone / analogs & derivatives
  • 4-Butyrolactone / metabolism
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Escherichia coli / cytology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Homoserine / analogs & derivatives
  • Homoserine / metabolism
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • Microscopy, Fluorescence
  • Promoter Regions, Genetic / genetics
  • Quorum Sensing*
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / methods*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Transformation, Genetic

Substances

  • Bacterial Proteins
  • LasI protein, Pseudomonas aeruginosa
  • LasR protein, Pseudomonas aeruginosa
  • N-(3-oxododecanoyl)homoserine lactone
  • Trans-Activators
  • Green Fluorescent Proteins
  • Homoserine
  • 4-Butyrolactone