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. 1996 Mar;62(3):778–783. doi: 10.1128/aem.62.3.778-783.1996

Use of an ipb-lux Fusion To Study Regulation of the Isopropylbenzene Catabolism Operon of Pseudomonas putida RE204 and To Detect Hydrophobic Pollutants in the Environment

O V Selifonova, R W Eaton
PMCID: PMC1388794  PMID: 16535269

Abstract

A DNA segment involved in the regulation of the isopropylbenzene (cumene) catabolism operon (ipb) of plasmid pRE4 from Pseudomonas putida RE204 and the Vibrio fischeri luciferase genes, luxCDABE, were used to create an ipbRo/pA(prm1)-luxCDABE reporter fusion plasmid, pOS25. Escherichia coli HMS174(pOS25) produces light in the presence of inducers of the ipb operon. These inducers were shown to be hydrophobic compounds and to include monoalkylbenzenes, substituted benzenes and toluenes, some alkanes and cycloalkanes, chlorinated solvents, and naphthalenes. Complex hydrocarbon mixtures, such as gasoline, diesel fuel, jet fuels (JP-4 and JP-5), and creosote, were also inducers of ipb-lux. Bacteria carrying the ipb-lux reporter may be useful as bioindicators of hydrocarbon pollution in the environment and may be particularly valuable for examining the bioavailability of inducing pollutants.

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Selected References

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  1. Abril M. A., Michan C., Timmis K. N., Ramos J. L. Regulator and enzyme specificities of the TOL plasmid-encoded upper pathway for degradation of aromatic hydrocarbons and expansion of the substrate range of the pathway. J Bacteriol. 1989 Dec;171(12):6782–6790. doi: 10.1128/jb.171.12.6782-6790.1989. [DOI] [PMC free article] [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. Eaton R. W., Nitterauer J. D. Biotransformation of benzothiophene by isopropylbenzene-degrading bacteria. J Bacteriol. 1994 Jul;176(13):3992–4002. doi: 10.1128/jb.176.13.3992-4002.1994. [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. Eaton R. W., Timmis K. N. Characterization of a plasmid-specified pathway for catabolism of isopropylbenzene in Pseudomonas putida RE204. J Bacteriol. 1986 Oct;168(1):123–131. doi: 10.1128/jb.168.1.123-131.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heald S., Jenkins R. O. Trichloroethylene removal and oxidation toxicity mediated by toluene dioxygenase of Pseudomonas putida. Appl Environ Microbiol. 1994 Dec;60(12):4634–4637. doi: 10.1128/aem.60.12.4634-4637.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. McClay K., Streger S. H., Steffan R. J. Induction of toluene oxidation activity in Pseudomonas mendocina KR1 and Pseudomonas sp. strain ENVPC5 by chlorinated solvents and alkanes. Appl Environ Microbiol. 1995 Sep;61(9):3479–3481. doi: 10.1128/aem.61.9.3479-3481.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mueller J. G., Middaugh D. P., Lantz S. E., Chapman P. J. Biodegradation of creosote and pentachlorophenol in contaminated groundwater: chemical and biological assessment. Appl Environ Microbiol. 1991 May;57(5):1277–1285. doi: 10.1128/aem.57.5.1277-1285.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nikaido H., Vaara M. Molecular basis of bacterial outer membrane permeability. Microbiol Rev. 1985 Mar;49(1):1–32. doi: 10.1128/mr.49.1.1-32.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pavel H., Forsman M., Shingler V. An aromatic effector specificity mutant of the transcriptional regulator DmpR overcomes the growth constraints of Pseudomonas sp. strain CF600 on para-substituted methylphenols. J Bacteriol. 1994 Dec;176(24):7550–7557. doi: 10.1128/jb.176.24.7550-7557.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Selifonova O., Burlage R., Barkay T. Bioluminescent sensors for detection of bioavailable Hg(II) in the environment. Appl Environ Microbiol. 1993 Sep;59(9):3083–3090. doi: 10.1128/aem.59.9.3083-3090.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Shingler V., Moore T. Sensing of aromatic compounds by the DmpR transcriptional activator of phenol-catabolizing Pseudomonas sp. strain CF600. J Bacteriol. 1994 Mar;176(6):1555–1560. doi: 10.1128/jb.176.6.1555-1560.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sikkema J., de Bont J. A., Poolman B. Mechanisms of membrane toxicity of hydrocarbons. Microbiol Rev. 1995 Jun;59(2):201–222. doi: 10.1128/mr.59.2.201-222.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Van Dyk T. K., Majarian W. R., Konstantinov K. B., Young R. M., Dhurjati P. S., LaRossa R. A. Rapid and sensitive pollutant detection by induction of heat shock gene-bioluminescence gene fusions. Appl Environ Microbiol. 1994 May;60(5):1414–1420. doi: 10.1128/aem.60.5.1414-1420.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wang Y., Rawlings M., Gibson D. T., Labbé D., Bergeron H., Brousseau R., Lau P. C. Identification of a membrane protein and a truncated LysR-type regulator associated with the toluene degradation pathway in Pseudomonas putida F1. Mol Gen Genet. 1995 Mar 10;246(5):570–579. doi: 10.1007/BF00298963. [DOI] [PubMed] [Google Scholar]
  18. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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