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. 1993 Sep;59(9):3083–3090. doi: 10.1128/aem.59.9.3083-3090.1993

Bioluminescent sensors for detection of bioavailable Hg(II) in the environment.

O Selifonova 1, R Burlage 1, T Barkay 1
PMCID: PMC182410  PMID: 8215378

Abstract

Biosensors for the detection of pollutants in the environment can complement analytical methods by distinguishing bioavailable from inert, unavailable forms of contaminants. By using fusions of the well-understood Tn21 mercury resistance operon (mer) with promoterless luxCDABE from Vibrio fischeri, we have constructed and tested three biosensors for Hg(II). Bioluminescence specified by pRB28, carrying merRo/pT, by pOS14, mediating active transport of Hg(II), and by pOS15, containing an intact mer operon, was measured in rich and minimal media. The highest sensitivities were achieved in minimal medium and were 1, 0.5, and 25 nM Hg(II) for pRB28, pOS14, and pOS15, respectively. The utility of the biosensors in natural waters was demonstrated with freshwater, rain, and estuarine samples supplemented with Hg(II). mer-lux carried by pRB28 and pOS14 responded to Hg(II) in mercury-contaminated water samples collected from a freshwater pond. Semiquantitative analyses based on light emission in samples collected from the inlet (analytically determined total mercury, approximately 20 nM) and outlet (total mercury, approximately 7 nM) of the pond showed bioavailable mercury at approximately 20 and 1 to 2 nM, respectively. Thus, the biosensors described here semiquantitatively detect bioavailable inorganic mercury (at a nanomolar to micromolar concentration range) in contaminated waters.

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Selected References

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  1. Barrineau P., Gilbert P., Jackson W. J., Jones C. S., Summers A. O., Wisdom S. The DNA sequence of the mercury resistance operon of the IncFII plasmid NR1. J Mol Appl Genet. 1984;2(6):601–619. [PubMed] [Google Scholar]
  2. Campbell J. L., Richardson C. C., Studier F. W. Genetic recombination and complementation between bacteriophage T7 and cloned fragments of T7 DNA. Proc Natl Acad Sci U S A. 1978 May;75(5):2276–2280. doi: 10.1073/pnas.75.5.2276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Condee C. W., Summers A. O. A mer-lux transcriptional fusion for real-time examination of in vivo gene expression kinetics and promoter response to altered superhelicity. J Bacteriol. 1992 Dec;174(24):8094–8101. doi: 10.1128/jb.174.24.8094-8101.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eaton R. W., Ribbons D. W. Metabolism of dibutylphthalate and phthalate by Micrococcus sp. strain 12B. J Bacteriol. 1982 Jul;151(1):48–57. doi: 10.1128/jb.151.1.48-57.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Farrell R. E., Germida J. J., Huang P. M. Biotoxicity of mercury as influenced by mercury(II) speciation. Appl Environ Microbiol. 1990 Oct;56(10):3006–3016. doi: 10.1128/aem.56.10.3006-3016.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Foster T. J., Nakahara H., Weiss A. A., Silver S. Transposon A-generated mutations in the mercuric resistance genes of plasmid R100-1. J Bacteriol. 1979 Oct;140(1):167–181. doi: 10.1128/jb.140.1.167-181.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gambill B. D., Summers A. O. Synthesis and degradation of the mRNA of the Tn21 mer operon. J Mol Biol. 1992 May 20;225(2):251–259. doi: 10.1016/0022-2836(92)90919-b. [DOI] [PubMed] [Google Scholar]
  8. Gambill B. D., Summers A. O. Versatile mercury-resistant cloning and expression vectors. Gene. 1985;39(2-3):293–297. doi: 10.1016/0378-1119(85)90326-9. [DOI] [PubMed] [Google Scholar]
  9. Gilmour C. C., Henry E. A. Mercury methylation in aquatic systems affected by acid deposition. Environ Pollut. 1991;71(2-4):131–169. doi: 10.1016/0269-7491(91)90031-q. [DOI] [PubMed] [Google Scholar]
  10. Guida L., Saidi Z., Hughes M. N., Poole R. K. Aluminium toxicity and binding to Escherichia coli. Arch Microbiol. 1991;156(6):507–512. doi: 10.1007/BF00245400. [DOI] [PubMed] [Google Scholar]
  11. Hamlett N. V., Landale E. C., Davis B. H., Summers A. O. Roles of the Tn21 merT, merP, and merC gene products in mercury resistance and mercury binding. J Bacteriol. 1992 Oct;174(20):6377–6385. doi: 10.1128/jb.174.20.6377-6385.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  13. Hastings J. W., Potrikus C. J., Gupta S. C., Kurfürst M., Makemson J. C. Biochemistry and physiology of bioluminescent bacteria. Adv Microb Physiol. 1985;26:235–291. doi: 10.1016/s0065-2911(08)60398-7. [DOI] [PubMed] [Google Scholar]
  14. Heitzer A., Webb O. F., Thonnard J. E., Sayler G. S. Specific and quantitative assessment of naphthalene and salicylate bioavailability by using a bioluminescent catabolic reporter bacterium. Appl Environ Microbiol. 1992 Jun;58(6):1839–1846. doi: 10.1128/aem.58.6.1839-1846.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Helmann J. D., Ballard B. T., Walsh C. T. The MerR metalloregulatory protein binds mercuric ion as a tricoordinate, metal-bridged dimer. Science. 1990 Feb 23;247(4945):946–948. doi: 10.1126/science.2305262. [DOI] [PubMed] [Google Scholar]
  16. King J. M., Digrazia P. M., Applegate B., Burlage R., Sanseverino J., Dunbar P., Larimer F., Sayler G. S. Rapid, sensitive bioluminescent reporter technology for naphthalene exposure and biodegradation. Science. 1990 Aug 17;249(4970):778–781. doi: 10.1126/science.249.4970.778. [DOI] [PubMed] [Google Scholar]
  17. Lee I. W., Gambill B. D., Summers A. O. Translation of merD in Tn21. J Bacteriol. 1989 Apr;171(4):2222–2225. doi: 10.1128/jb.171.4.2222-2225.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meighen E. A. Molecular biology of bacterial bioluminescence. Microbiol Rev. 1991 Mar;55(1):123–142. doi: 10.1128/mr.55.1.123-142.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakahara H., Silver S., Miki T., Rownd R. H. Hypersensitivity to Hg2+ and hyperbinding activity associated with cloned fragments of the mercurial resistance operon of plasmid NR1. J Bacteriol. 1979 Oct;140(1):161–166. doi: 10.1128/jb.140.1.161-166.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nucifora G., Chu L., Silver S., Misra T. K. Mercury operon regulation by the merR gene of the organomercurial resistance system of plasmid pDU1358. J Bacteriol. 1989 Aug;171(8):4241–4247. doi: 10.1128/jb.171.8.4241-4247.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nucifora G., Silver S., Misra T. K. Down regulation of the mercury resistance operon by the most promoter-distal gene merD. Mol Gen Genet. 1989 Dec;220(1):69–72. doi: 10.1007/BF00260858. [DOI] [PubMed] [Google Scholar]
  22. Park S. J., Wireman J., Summers A. O. Genetic analysis of the Tn21 mer operator-promoter. J Bacteriol. 1992 Apr;174(7):2160–2171. doi: 10.1128/jb.174.7.2160-2171.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ralston D. M., O'Halloran T. V. Ultrasensitivity and heavy-metal selectivity of the allosterically modulated MerR transcription complex. Proc Natl Acad Sci U S A. 1990 May;87(10):3846–3850. doi: 10.1073/pnas.87.10.3846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rogowsky P. M., Close T. J., Chimera J. A., Shaw J. J., Kado C. I. Regulation of the vir genes of Agrobacterium tumefaciens plasmid pTiC58. J Bacteriol. 1987 Nov;169(11):5101–5112. doi: 10.1128/jb.169.11.5101-5112.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ross W., Park S. J., Summers A. O. Genetic analysis of transcriptional activation and repression in the Tn21 mer operon. J Bacteriol. 1989 Jul;171(7):4009–4018. doi: 10.1128/jb.171.7.4009-4018.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Silver S., Walderhaug M. Gene regulation of plasmid- and chromosome-determined inorganic ion transport in bacteria. Microbiol Rev. 1992 Mar;56(1):195–228. doi: 10.1128/mr.56.1.195-228.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Summers A. O. Organization, expression, and evolution of genes for mercury resistance. Annu Rev Microbiol. 1986;40:607–634. doi: 10.1146/annurev.mi.40.100186.003135. [DOI] [PubMed] [Google Scholar]
  28. Summers A. O. Untwist and shout: a heavy metal-responsive transcriptional regulator. J Bacteriol. 1992 May;174(10):3097–3101. doi: 10.1128/jb.174.10.3097-3101.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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