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*

*Corresponding author for this work
    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 publicationc-di-GMP Signaling : Methods and Protocols
    EditorsKarin Sauer
    Number of pages12
    Volume1657
    PublisherHumana Press
    Publication dateJan 2017
    Pages87-98
    ISBN (Print)978-1-4939-7239-5
    ISBN (Electronic)978-1-4939-7240-1
    DOIs
    Publication statusPublished - Jan 2017
    SeriesMethods in Molecular Biology
    Volume1657
    ISSN1064-3745

    Keywords

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

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