Skip to main content

Detection of Oxidants Using lux Fusions to Oxidative Stress Promoters

  • Protocol
Bioluminescence Methods and Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 102))

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 (25) and will not be detailed here.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. 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.

    Google Scholar 

  2. 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

    Google Scholar 

  3. 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

    Google Scholar 

  4. 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.

    CAS  Google Scholar 

  5. Demple, B. 1991. Regulation of bacterial oxidative stress genes. Ann Rev. Genet 25, 315–337.

    Article  PubMed  CAS  Google Scholar 

  6. 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.

    Google Scholar 

  7. 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

    Article  CAS  Google Scholar 

  8. 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

    CAS  Google Scholar 

  9. 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.

    Google Scholar 

  10. 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.

    Article  CAS  Google Scholar 

  11. 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

    Article  CAS  Google Scholar 

  12. 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

    Google Scholar 

  13. 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.

    Article  CAS  Google Scholar 

  14. Miller, J. H. (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  15. 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

    Google Scholar 

  16. 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.

    Google Scholar 

  17. 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

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

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

Download citation

  • DOI: https://doi.org/10.1385/0-89603-520-4:189

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-520-1

  • Online ISBN: 978-1-59259-577-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics