1. Improved bacterial SOS promoter∷lux fusions for genotoxicity detection.
- Author
-
Davidov Y, Rozen R, Smulski DR, Van Dyk TK, Vollmer AC, Elsemore DA, LaRossa RA, and Belkin S
- Subjects
- Alginates, Bacteria drug effects, Bacterial Outer Membrane Proteins genetics, Bacterial Proteins genetics, Bacterial Proteins metabolism, Escherichia coli genetics, Escherichia coli Proteins, Genes, Reporter genetics, Glucuronic Acid, Hexuronic Acids, Hydrogen Peroxide toxicity, Industrial Waste, Kinetics, Luminescent Measurements, Membrane Transport Proteins, Mutation, Photorhabdus genetics, Recombinant Fusion Proteins drug effects, Recombinant Fusion Proteins genetics, Replicon, SOS Response, Genetics drug effects, Salmonella typhimurium genetics, Serine Endopeptidases genetics, Serine Endopeptidases metabolism, Species Specificity, Temperature, Time Factors, Vibrio genetics, Water Pollutants toxicity, Bacteria genetics, Mutagenicity Tests methods, Promoter Regions, Genetic genetics, Repressor Proteins genetics, SOS Response, Genetics genetics, Trans-Activators genetics
- Abstract
Escherichia coli strains containing plasmid-borne fusions of Vibrio fischeri lux to the recA promoter-operator region were previously shown to be potentially useful for detecting genotoxicants. In an attempt to improve past performance, the present study examines several modifications and variations of this design, singly or in various combinations: (1) modifying the host cell's toxicant efflux capacity via a tolC mutation; (2) incorporating the lux fusion onto the bacterial chromosome, rather then on a plasmid; (3) changing the reporter element to a different lux system (Photorhabdus luminescens), with a broader temperature range; (4) using Salmonella typhimurium instead of an E. coli host. A broad spectrum of responses to pure chemicals as well as to industrial wastewater samples was observed. Generally, fastest responses were exhibited by Sal94, a S. typhimurium strain harboring a plasmid-borne fusion of V. fischeri lux to the E. coli recA promoter. Highest sensitivity, however, was demonstrated by DPD3063, an E. coli strain in which the same fusion was integrated into the bacterial chromosome, and by DPD2797, a plasmid-bearing tolC mutant. Overall, the two latter strains appeared to perform better and seemed preferable over the others. The sensor strains retained their sensitivity following a 2-month incubation after alginate-embedding, but at the cost of a significantly delayed response.
- Published
- 2000
- Full Text
- View/download PDF