Abstract
This chapter provides basic instructions in the use of genetically engineered Escherichia coli strains that luminesce in response to the presence of oxidants. These strains carry plasmid dertvatives of pUCD615 (1) in which Vibrio fischeri 1uxCDABE is driven by selected promoters of genes responstve to oxidative stress, The construction of such plasmids has been reported before (2–5) and will not be detailed here.
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References
Rogowsky, P. M., Close, T. J., Chimera, J. A., Shaw, J. J., and Kado, C. I. (1987) Regulation of the vir genes of Agrobacterium tumefaciens plasmid pTiC58. J Bacterzol. 169, 510l–5112.
Van Dyk, T. K, Majartan, W. R., Konstantinov, K. B., Young, R. M., Dhuqati, P S., and LaRossa, R. A. (1994) Rapid and sensitive pollutant detection by induction of heat shock gene-biolummescence gene fusions. Appl Environ. Microbial 60, 1414–1420
Van Dyk, T K, Smulski, D. R, Reed, T R., Belkin, S, Vollmer, A C, and LaRossa, R A. (1995) Responses to toxicants of an Escherichia coli stram carrying a uspA ′·· lux genetic fusion and an E coli strain carrying a grpE′·· lux fusion are similar. Appl Envzron Microbzol. 61, 4124–4127
Belkin, S., Smulski, D. R., Vollmer, A. C, Van Dyk, T. K, and LaRossa, R. A. (1996) Oxidative stress detection with Escherzchza colz bearmg a katG′ lux fusion. Appl Envzron Microbial 62, 2252–2256.
Demple, B. 1991. Regulation of bacterial oxidative stress genes. Ann Rev. Genet 25, 315–337.
Netdhardt, F. C. and VanBogelen, R. H. (1987) Heat shock response, in Escherichia colz and Salmonella typhimurium Cellular and Molecular Biology (Netdhardt, F. C, Ingraham, J. L., Low, K. B., Magasamk, B., Schaechter, M., and Umbarger, H. E, eds.), ASM, Washington, DC, pp. 1334–1345.
Lange, R. and Hengge-Aronis, R. (1991) Identification of a central regulator of stationary-phase gene expression in Escherzchza coli Mol Microbiol. 5, 49–59
Greenberg, J. T., Chou, J. H., Monach, P. A., and Demple, B. (1991) Activation of oxidative stress genes by mutations at the soxQ/cfxB/marA locus of Escherzchza colz J.Bacterzol. 173, 4433–4439
Compan, I. and Touati, D. (1993) Interaction of six global transcription regulators in expression of manganese superoxide dismutase in Escherichia coli K-12. J Bacterzol. 175, 1687–1696.
Stanley, P. E. (1996) Commercially available luminometers and imaging devtces for low-light level measurements and kits and reagents utilizing bioluminescence or chemi-luminescence: survey update 4. J Biolumznescence Chemiluminescence 11, 175–191.
Menzel, R. (1989) A microtiter plate-based system for the semiautomated growth and assay of bacterial cells for β-galactosidase activity Anal. Bzochem 181, 40–50
Belkm, S., Vollmer, A. C., Van Dyk, T. K, Smulski, D. R., Reed, T. R., and LaRossa, R. A. (1994) Oxidative and DNA damaging agents induce luminescence in E coli harbormg lux fusions to stress promoters, in Bioluminescence and Chemiluminescence Fundamentals and Applzed Aspects (Campbell, A. K, Kricka, L. J., and Stanley, P. E, eds.), John Wiley, Chichester, pp 509–512
Belkin, S., Van Dyk, T. K., Vollmer, A. C., Smulski, D. R., and LaRossa, R A. (1996). Monitoring sub-toxic environmental hazards by stress-responsive luminous bacteria Environ Toxicol Water Quality 11, 179–185.
Miller, J. H. (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Belkm, S., Smulski, D. R., Dadon, S., Vollmer, A C, Van Dyk, T. K., and LaRossa, R. A. A panel of stress-responsive lummous bacteria for the detection of specific classes of toxicants. Water Res., in press
Belkibin, S Stress responsive luminous bacteria for toxicity and genotoxicity momtoring, in Microscale Aquatic Toxicology—Advances, Techniques and Practice (Wells, P. G., Lee, K., and Blalse, C, eds.), CRC Lewis, Florida, in press.
Dukan, S., Dadon, S., Smulski, D. R., and Belkm, S. (1996) Hypochlorous acid activates the heat shock and soxRS systems of Escherichia co1i Appl. Environ. Microbial 62, 4003–4008
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© 1998 Humana Press Inc., Totowa, NJ
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Belkin, S. (1998). Detection of Oxidants Using lux Fusions to Oxidative Stress Promoters. In: LaRossa, R.A. (eds) Bioluminescence Methods and Protocols. Methods in Molecular Biology™, vol 102. Humana Press. https://doi.org/10.1385/0-89603-520-4:189
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DOI: https://doi.org/10.1385/0-89603-520-4:189
Publisher Name: Humana Press
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