Detection of N-acyl homoserine lactones using a traI-luxCDABE-based biosensor as a high-throughput screening tool.
BACKGROUND
Bacteria use N-acyl homoserine lactone (AHL) molecules to regulate the expression of genes in a density-dependent manner. Several biosensors have been developed and engineered to detect the presence of all types of AHLs.


RESULTS
In this study, we describe the usefulness of a traI-luxCDABE-based biosensor to quickly detect AHLs from previously characterized mutants of Burkholderia cenocepacia and Pseudomonas aeruginosa in both liquid and soft-agar co-culture assays in a high-throughput manner. The technique uses a co-culture system where the strain producing the AHLs is grown simultaneously with the reporter strain. Use of this assay in liquid co-culture allows the measurement of AHL activity in real time over growth. We tested this assay with Burkholderia cenocepacia and Pseudomonas aeruginosa but it should be applicable to a broad range of gram negative species that produce AHLs.


CONCLUSIONS
The co-culture assays described enable the detection of AHL production in both P. aeruginosa and B. cenocepacia and should be applicable to AHL analysis in other bacterial species. The high-throughput adaptation of the liquid co-culture assay could facilitate the screening of large libraries for the identification of mutants or compounds that block the synthesis or activity of AHLs.
Bernier
Steve P
SP
Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Canada. sbernier@pasteur.fr
Beeston
Anne L
AL
Sokol
Pamela A
PA
D054742:Acyl-Butyrolactones / Q000032:analysis*; D016960:Agrobacterium tumefaciens / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D015374:Biosensing Techniques; D016956:Burkholderia cepacia / Q000378:metabolism*; D005798:Genes, Bacterial; D017930:Genes, Reporter; D008163:Luminescent Measurements; D009876:Operon; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D013997:Time Factors
D054742:Acyl-Butyrolactones; D001426:Bacterial Proteins; C080512:TraR protein, Agrobacterium tumefaciens