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Applied and Environmental Microbiology, February 2001, p. 575-585, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.575-585.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

gfp-Based N-Acyl Homoserine-Lactone Sensor Systems for Detection of Bacterial Communication

Jens Bo Andersen,1 Arne Heydorn,1 Morten Hentzer,1 Leo Eberl,2 Otto Geisenberger,2 Bjarke Bak Christensen,1,dagger Søren Molin,1 and Michael Givskov1,*

Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark,1 and Lehrstuhl für Mikrobiologie, Technische Universität München, D-80290 Munich, Germany2

Received 19 June 2000/Accepted 9 November 2000

In order to perform single-cell analysis and online studies of N-acyl homoserine lactone (AHL)-mediated communication among bacteria, components of the Vibrio fischeri quorum sensor encoded by luxR-PluxI have been fused to modified versions of gfpmut3* genes encoding unstable green fluorescent proteins. Bacterial strains harboring this green fluorescent sensor detected a broad spectrum of AHL molecules and were capable of sensing the presence of 5 nM N-3-oxohexanoyl-L-homoserine lactone in the surroundings. In combination with epifluorescent microscopy, the sensitivity of the sensor enabled AHL detection at the single-cell level and allowed for real-time measurements of fluctuations in AHL concentrations. This green fluorescent AHL sensor provides a state-of-the-art tool for studies of communication between the individuals present in mixed bacterial communities.

* Corresponding author. Mailing address: Department of Microbiology, Building 301, The Technical University of Denmark, DK-2800 Lyngby, Denmark. Phone: 45 45 25 27 69. Fax: 45 45 93 28 09. E-mail: immg{at}pop.dtu.dk.

dagger Present address: The Danish Veterinary and Food Administration, 2860 Søborg, Denmark.

Applied and Environmental Microbiology, February 2001, p. 575-585, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.575-585.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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