Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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4-Butyrolactone / analogs & derivatives*
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4-Butyrolactone / genetics
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4-Butyrolactone / metabolism*
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Biosensing Techniques / methods*
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Chemotaxis / physiology
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Gene Expression Regulation, Bacterial
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Gram-Negative Bacteria / genetics
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Gram-Negative Bacteria / physiology*
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Green Fluorescent Proteins
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Image Processing, Computer-Assisted / methods
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism*
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Microscopy, Confocal
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Plasmids / genetics
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Promoter Regions, Genetic / genetics
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Repressor Proteins / genetics
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Repressor Proteins / metabolism
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Signal Transduction*
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Trans-Activators / genetics
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Trans-Activators / metabolism
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Transcription Factors / genetics
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Transcription Factors / metabolism
Substances
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Bacterial Proteins
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Luminescent Proteins
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LuxI protein, Bacteria
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Repressor Proteins
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Trans-Activators
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Transcription Factors
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LuxR autoinducer binding proteins
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homoserine lactone
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Green Fluorescent Proteins
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N-(3-oxohexanoyl)-3-aminodihydro-2(3H)-furanone
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4-Butyrolactone