Gauging and visualizing c-di-GMP levels in pseudomonas aeruginosa using fluorescence-based biosensors

Morten Rybtke, Song Lin Chua, Joey Kuok Hoong Yam, Michael Givskov, Liang Yang, Tim Tolker-Nielsen

Research output: Chapter in book / Conference proceedingChapter in an edited book (as author)Academic researchpeer-review

2 Citations (Scopus)

Abstract

Recent research has shown that the molecule c-di-GMP is an important second messenger regulating various functions in bacteria. In particular, the implication of c-di-GMP as a positive regulator of adhesion and biofilm formation has gained momentum as a highly relevant research topic, as detailed knowledge about the underlying regulatory mechanisms may enable the development of measures to control biofilms in both industrial and medical settings. Accordingly, it is in many cases of interest to measure the c-di-GMP level in bacteria under specific conditions or in specific mutant strains. We have developed a collection of fluorescence-based c-di-GMP biosensors capable of gauging the c-di-GMP level in Pseudomonas aeruginosa and closely related bacteria. Here, we describe protocols for the use of these biosensors in gauging and visualizing cellular c-di-GMP levels of P. aeruginosa both in in vitro setups such as continuous-culture flow-cell biofilms, and in in vivo settings such as a murine corneal infection model.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages87-98
Number of pages12
DOIs
Publication statusPublished - 10 Sep 2017
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1657
ISSN (Print)1064-3745

Keywords

  • Biofilm
  • Biosensor
  • c-di-GMP
  • Cyclic di-GMP
  • Fluorescence
  • Ocular infection
  • Pseudomonas

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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