Enhancement of the multi-channel continuous monitoring system through the use of Xenorhabdus luminescens lux fusions.
The enhancement of the multi-channel continuous toxicity monitoring system developed previously was studied. To achieve better and more stable results from the system, the use of thermo-lux fusion strains that express the luxCDABE genes from Xenorhabdus luminescens was evaluated. A total of six recombinant Escherichia coli strains with the promoters from three oxidative-stress responsive genes, i.e. the katG, sodA and pqi-5 genes, fused to either the lux genes from Vibrio fischeri or X. luminescens were characterized and their responses to different chemicals compared. It was found that the basal level bioluminescence (BL) from the thermo-lux fusion strains was always higher while that of the V. fischeri lux strains were always near or below the lower limit of detection of the system. For example, the katG::V. fischeri lux strain, DPD2511, gave no discernible response due to its low level expression while a fusion of the katG promoter with the X. luminescens lux operon was clearly responsive and capable of detecting hydrogen peroxide down to about 1 ppm. The use of the thermo-lux strains found them to be as sensitive as the V. fischeri lux strains while providing a brighter, more stable basal level bioluminescence, making the analysis and monitoring of water-borne toxicity more reliable.
Lee
Jin Hyung
JH
National Research Laboratory on Environmental Biotechnology, Department of Environmental Science and Engineering, Kwangju Institute of Science and Technology (K-JIST), 1 Oryong-dong, Puk-gu, Gwangju 500-712, South Korea.
Mitchell
Robert J
RJ
Gu
Man Bock
MB
D048908:Aliivibrio fischeri / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D018920:Coculture Techniques / Q000295:instrumentation / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000295:instrumentation / Q000379:methods; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D024861:Genetic Enhancement / Q000379:methods; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009862:Online Systems; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D020637:Photorhabdus / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014874:Water Pollutants, Chemical / Q000032:analysis
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D014874:Water Pollutants, Chemical; D006861:Hydrogen Peroxide