gfp-based N-acyl homoserine-lactone sensor systems for detection of bacterial communication.
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-P(luxI) 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.
Andersen
J B
JB
Department of Microbiology, The Technical University of Denmark, DK-2800 Lyngby, Denmark.
Heydorn
A
A
Hentzer
M
M
Eberl
L
L
Geisenberger
O
O
Christensen
B B
BB
Molin
S
S
Givskov
M
M
D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D002633:Chemotaxis / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000502:physiology*; D049452:Green Fluorescent Proteins; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D008164:Luminescent Proteins; C085427:LuxI protein, Bacteria; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C064778:LuxR autoinducer binding proteins; C088386:homoserine lactone; D049452:Green Fluorescent Proteins; C030654:N-(3-oxohexanoyl)-3-aminodihydro-2(3H)-furanone; D015107:4-Butyrolactone