Cyclic di-GMP-Responsive Transcriptional Reporter Bioassays in Pseudomonas aeruginosa

Methods Mol Biol. 2017:1657:99-110. doi: 10.1007/978-1-4939-7240-1_9.

Abstract

3',5'-cyclic diguanosine monophosphate (cyclic di-GMP) is a bacterial secondary messenger molecule that regulates many important cellular activities and behaviors, such as motility and biofilm formation. While mass spectrometry protocols for quantitative analyses of intracellular cyclic di-GMP concentrations have been developed, they are time intensive, expensive, low-throughput, and incapable of directly monitoring dynamic changes in vivo. In this protocol, we provide a Pseudomonas aeruginosa-specific detailed methodology to assay the intracellular levels of cyclic di-GMP using biological reporters.

Keywords: Cyclic di-GMP; Fluorescence assay; Luminescence assay; Pseudomonas aeruginosa; c-di-GMP.

MeSH terms

  • Cyclic GMP / analogs & derivatives*
  • Cyclic GMP / metabolism
  • Gene Expression*
  • Genes, Reporter*
  • Luminescent Measurements
  • Microscopy, Fluorescence
  • Pseudomonas aeruginosa / genetics*
  • Pseudomonas aeruginosa / metabolism*
  • Second Messenger Systems*
  • Transcription, Genetic*

Substances

  • bis(3',5')-cyclic diguanylic acid
  • Cyclic GMP