Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa

Publication: Research - peer-reviewJournal article – Annual report year: 2012

Standard

Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa. / Rybtke, Morten T.; Borlee, Bradley R.; Murakami, Keiji; Irie, Yasuhiko; Hentzer, Morten; Nielsen, Thomas Eiland; Givskov, Michael; Parsek, Matthew R.; Tolker-Nielsen, Tim.

In: Applied and Environmental Microbiology, Vol. 78, No. 15, 2012, p. 5060-5069.

Publication: Research - peer-reviewJournal article – Annual report year: 2012

Harvard

Rybtke, MT, Borlee, BR, Murakami, K, Irie, Y, Hentzer, M, Nielsen, TE, Givskov, M, Parsek, MR & Tolker-Nielsen, T 2012, 'Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa' Applied and Environmental Microbiology, vol 78, no. 15, pp. 5060-5069., 10.1128/AEM.00414-12

APA

Rybtke, M. T., Borlee, B. R., Murakami, K., Irie, Y., Hentzer, M., Nielsen, T. E., ... Tolker-Nielsen, T. (2012). Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa. Applied and Environmental Microbiology, 78(15), 5060-5069. 10.1128/AEM.00414-12

CBE

Rybtke MT, Borlee BR, Murakami K, Irie Y, Hentzer M, Nielsen TE, Givskov M, Parsek MR, Tolker-Nielsen T. 2012. Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa. Applied and Environmental Microbiology. 78(15):5060-5069. Available from: 10.1128/AEM.00414-12

MLA

Vancouver

Author

Rybtke, Morten T.; Borlee, Bradley R.; Murakami, Keiji; Irie, Yasuhiko; Hentzer, Morten; Nielsen, Thomas Eiland; Givskov, Michael; Parsek, Matthew R.; Tolker-Nielsen, Tim / Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa.

In: Applied and Environmental Microbiology, Vol. 78, No. 15, 2012, p. 5060-5069.

Publication: Research - peer-reviewJournal article – Annual report year: 2012

Bibtex

@article{657f12beaca24f328c07ecce70e1eda9,
title = "Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa",
publisher = "American Society for Microbiology",
author = "Rybtke, {Morten T.} and Borlee, {Bradley R.} and Keiji Murakami and Yasuhiko Irie and Morten Hentzer and Nielsen, {Thomas Eiland} and Michael Givskov and Parsek, {Matthew R.} and Tim Tolker-Nielsen",
year = "2012",
doi = "10.1128/AEM.00414-12",
volume = "78",
number = "15",
pages = "5060--5069",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",

}

RIS

TY - JOUR

T1 - Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa

A1 - Rybtke,Morten T.

A1 - Borlee,Bradley R.

A1 - Murakami,Keiji

A1 - Irie,Yasuhiko

A1 - Hentzer,Morten

A1 - Nielsen,Thomas Eiland

A1 - Givskov,Michael

A1 - Parsek,Matthew R.

A1 - Tolker-Nielsen,Tim

AU - Rybtke,Morten T.

AU - Borlee,Bradley R.

AU - Murakami,Keiji

AU - Irie,Yasuhiko

AU - Hentzer,Morten

AU - Nielsen,Thomas Eiland

AU - Givskov,Michael

AU - Parsek,Matthew R.

AU - Tolker-Nielsen,Tim

PB - American Society for Microbiology

PY - 2012

Y1 - 2012

N2 - The increased tolerance toward the host immune system and antibiotics displayed by biofilm-forming Pseudomonas aeruginosa and other bacteria in chronic infections such as cystic fibrosis bronchopneumonia is of major concern. Targeting of biofilm formation is believed to be a key aspect in the development of novel antipathogenic drugs that can augment the effect of classic antibiotics by decreasing antimicrobial tolerance. The second messenger cyclic di-GMP is a positive regulator of biofilm formation, and cyclic di-GMP signaling is now regarded as a potential target for the development of antipathogenic compounds. Here we describe the development of fluorescent monitors that can gauge the cellular level of cyclic di-GMP in P. aeruginosa. We have created cyclic di-GMP level reporters by transcriptionally fusing the cyclic di-GMP-responsive cdrA promoter to genes encoding green fluorescent protein. We show that the reporter constructs give a fluorescent readout of the intracellular level of cyclic di-GMP in P. aeruginosa strains with different levels of cyclic di-GMP. Furthermore, we show that the reporters are able to detect increased turnover of cyclic di-GMP mediated by treatment of P. aeruginosa with the phosphodiesterase inducer nitric oxide. Considering that biofilm formation is a necessity for the subsequent development of a chronic infection and therefore a pathogenicity trait, the reporters display a significant potential for use in the identification of novel antipathogenic compounds targeting cyclic di-GMP signaling, as well as for use in research aiming at understanding the biofilm biology of P. aeruginosa.

AB - The increased tolerance toward the host immune system and antibiotics displayed by biofilm-forming Pseudomonas aeruginosa and other bacteria in chronic infections such as cystic fibrosis bronchopneumonia is of major concern. Targeting of biofilm formation is believed to be a key aspect in the development of novel antipathogenic drugs that can augment the effect of classic antibiotics by decreasing antimicrobial tolerance. The second messenger cyclic di-GMP is a positive regulator of biofilm formation, and cyclic di-GMP signaling is now regarded as a potential target for the development of antipathogenic compounds. Here we describe the development of fluorescent monitors that can gauge the cellular level of cyclic di-GMP in P. aeruginosa. We have created cyclic di-GMP level reporters by transcriptionally fusing the cyclic di-GMP-responsive cdrA promoter to genes encoding green fluorescent protein. We show that the reporter constructs give a fluorescent readout of the intracellular level of cyclic di-GMP in P. aeruginosa strains with different levels of cyclic di-GMP. Furthermore, we show that the reporters are able to detect increased turnover of cyclic di-GMP mediated by treatment of P. aeruginosa with the phosphodiesterase inducer nitric oxide. Considering that biofilm formation is a necessity for the subsequent development of a chronic infection and therefore a pathogenicity trait, the reporters display a significant potential for use in the identification of novel antipathogenic compounds targeting cyclic di-GMP signaling, as well as for use in research aiming at understanding the biofilm biology of P. aeruginosa.

U2 - 10.1128/AEM.00414-12

DO - 10.1128/AEM.00414-12

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 15

VL - 78

SP - 5060

EP - 5069

ER -