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PMID: 10751731 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Improved bacterial SOS promoter∷lux fusions for genotoxicity detection.

Mutation research ·Vol. 466 ·No. 1 ·2000-03-03 ·Pages 97-107

Davidov Y, Rozen R, Smulski DR, Van Dyk TK, Vollmer AC, Elsemore DA, LaRossa RA, Belkin S

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.

MeSH Terms
Alginates Bacteria/drug effects,genetics Bacterial Outer Membrane Proteins/genetics Bacterial Proteins/genetics,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 Mutagenicity Tests/methods Mutation Photorhabdus/genetics Promoter Regions, Genetic/genetics Recombinant Fusion Proteins/drug effects,genetics Replicon Repressor Proteins/genetics SOS Response, Genetics/drug effects,genetics Salmonella typhimurium/genetics Serine Endopeptidases/genetics,metabolism Species Specificity Temperature Time Factors Trans-Activators/genetics Vibrio/genetics Water Pollutants/toxicity
Chemicals
Alginates Bacterial Outer Membrane Proteins Bacterial Proteins Escherichia coli Proteins Hexuronic Acids Industrial Waste LexA protein, Bacteria Membrane Transport Proteins Recombinant Fusion Proteins Repressor Proteins Trans-Activators Water Pollutants tolC protein, E coli LuxR autoinducer binding proteins Glucuronic Acid Hydrogen Peroxide Serine Endopeptidases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Davidov Y
Division of Environmental Sciences, the Fredy and Nadine Herrmann Graduate School of Applied Science, The Hebrew University of Jerusalem, Jerusalem, Israel.
Rozen R
Smulski D R
Van Dyk T K
Vollmer A C
Elsemore D A
LaRossa R A
Belkin S
Article Info
Journal
Mutation research
Abbr.
Mutat Res
ISSN
0027-5107
Published
2000-03-03
Pages
97-107
Language
English
Region
Netherlands
NLM ID
0400763
Subset
IM
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