Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/100527
Title: Fluorescence-based reporter for gauging cyclic Di-GMP levels in Pseudomonas aeruginosa
Authors: Borlee, Bradley R.
Nielsen, Thomas E.
Parsek, Matthew R.
Rybtke, Morten Theil
Murakami, Keiji
Irie, Yasuhiko
Hentzer, Morten
Givskov, Michael
Tolker-Nielsen, Tim
Issue Date: 2012
Source: Rybtke, M. T., Borlee, B. R., Murakami, K., Irie, Y., Hentzer, M., Nielsen, T. E., et al. (2012). Fluorescence-based reporter for gauging cyclic Di-GMP levels in Pseudomonas aeruginosa. Applied and Environmental Microbiology, 78(15), 5060-5069.
Series/Report no.: Applied and environmental microbiology
Abstract: 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.
URI: https://hdl.handle.net/10356/100527
http://hdl.handle.net/10220/10971
ISSN: 0099-2240
DOI: 10.1128/AEM.00414-12
Rights: © 2012 American Society for Microbiology.
Fulltext Permission: none
Fulltext Availability: No Fulltext
Appears in Collections:SCELSE Journal Articles

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