title: Detection and characterization of N-acyl-l-homoserine lactones using GFP-based biosensors in conjunction with thin-layer chromatography.
abstract: Many microorganisms use quorum sensing to regulate several complex phenotypes, and this is accomplished by the release of a signal molecule(s) into the environment. N-acyl-homoserine lactones (AHLs) are a common class of signalling molecule utilized by a range of microorganisms (primarily Gram negative bacteria but most recently also archaea) and are often detected through the use of bacterial biosensors. Biosensors can be limited by both their specificity and sensitivity, and the aim of this study was to modify and improve current AHL detection strategies. The biosensor employed in the present study was Escherichia coli MT102 harbouring a plasmid containing a LuxR based biosensor, which produces green fluorescent protein (GFP) as a reporting mechanism. A new method of visualizing the GFP based biosensor overlaid on silica sheets for the purpose of thin-layer chromatography (TLC) is presented. This new method vastly improves sensitivity of AHL detection by a GFP biosensor than previously reported and as such represents a powerful new tool in AHL research.
authors.affiliation: School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia; Australian Centre for Astrobiology, University of New South Wales Sydney, Australia.
authors.forename: James
authors.initials: J
authors.lastname: Charlesworth
authors.affiliation: School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia.
authors.forename: Onder
authors.initials: O
authors.lastname: Kimyon
authors.affiliation: School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia.
authors.forename: Michael
authors.initials: M
authors.lastname: Manefield
authors.affiliation: School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia; Australian Centre for Astrobiology, University of New South Wales Sydney, Australia. Electronic address: brendan.burns@unsw.edu.au.
authors.forename: Brendan P
authors.initials: BP
authors.lastname: Burns
mesh_terms: D054742:Acyl-Butyrolactones / Q000032:analysis*; D015374:Biosensing Techniques / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D012680:Sensitivity and Specificity
chemical_list: D054742:Acyl-Butyrolactones; D049452:Green Fluorescent Proteins
keywords: AHL (N-acyl-homoserine lactone); Biosensor; GFP (green fluorescent protein); Intercellular signalling; TLC (thin-layer chromatography)