Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-06-06
1998-07-28
Robinson, Douglas W.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 6, 435 8, 4351723, 43525233, 4353201, 435822, 435829, 435909, 536 232, 536 237, 536 231, 536 241, 935 72, 935 73, C12N 121, C12N 1563, C12Q 166, C07H 2104
Patent
active
057861625
DESCRIPTION:
BRIEF SUMMARY
The invention relates to fused genes, vectors containing them, process for preparing them and their use for determining the presence of metals or of xenobiotic compounds.
Toxical wastes are a significant contamination problem for a range of industries.
Among the substances involved, one may cite heavy metals and xenobiotic compounds which are very polluting and which may endanger health. The sources of pollution are varied. Moreover, with the enforcement of strict regulations, in order to limit the wastes containing metals such as heavy metals and xenobiotic compounds, there is a need for methods of detection of metals and xenobiotic compounds in environment.
Most of the methods used routinely to measure metal concentrations are physical methods which rely on the substantial physical (usually electronic) differences between the metal and the carrier medium.
Among these methods, the most commonly used are the inductively coupled plasma systems, the X-ray fluorescence or the atomic absorption.
The main advantage of these methods is the very low limits of detection (about 0,1 ppm) as well as a multielementary aspect of the analysis. But, the main drawbacks are the high price of the equipment, the use of which requires high qualified people, the long time required for preparing the samples to be analyzed and the sensitivity of these methods related to many interferences due to the nature of the samples.
Many organisms can tolerate high concentrations of heavy metals such as cadmium and lead. The mechanism involved varies. Specific, genetically coded resistance to heavy metals can evolve in populations of organisms exposed over long periods of time to heavy metals ("Genetic adaptation to heavy metals in aquatic organisms: a review" P. L. Klerks, J. S. Weis (1987), Environmental pollution 45: 173-205). Searches of soil in sites heavily contaminated with heavy metals routinely reveal strains of microorganisms with enhanced abilities to tolerate heavy metals. Several such strains have been isolated from a heavily contaminated site in Belgium, and the genetics of their responses to heavy metals have been analyzed ("Alcaligenes eutrophus CH34 is a facultative chemilithotroph with plasmid-bound resistance to heavy metals" M. Mergeay, D. Nies, H. G. Schlegel, J. Gerits, P. Charles, F. van Gijsegem (1985), J. Bacteriol. 162: 328-334; "Cloning of plasmid genes encoding resistance to cadmium, zinc, and cobalt in Alcaligenes eutrophus CH34" D. Nies, M. Mergeay, B. Friedrich, H. G. Schlegel (1987), J. Bacteriol. 169:4865-4868).
Most microorganisms can degrade a wide variety of compounds to generate metabolic energy and to make available metabolic intermediates, and particularly carbon, for their use. Some organisms specialize in the degradation of exotic materials, using unusual enzyme systems to do so. These are frequently soil bacteria that have evolved in sites where industrial activity has released a substantial amount of such material into the soil. The ability to degrade highly conjugated aromatic hydrocarbons and their halide derivatives is a good example, as these materials are rarely found in nature and require special enzymes to initiate their degradation, usually by oxygenation.
Alcaligenes eutrophus, as a bacterial organism, presents specific inducible genes of resistance with respect to heavy metals or involved in the catabolism of xenobiotics such as PCBs.
The bacteria of the group of Alcaligenes eutrophus (gram negative) have beside the property of being facultative chemilithotroph, the property of comprising one of several megaplasmids which confer on them multiple resistances with respect to heavy metals. These bacteria have been discovered in the neighborhood of non ferrous metal facturies and in the neighborhood of mining sites in Belgium and in Zaire (Diels et al., 1988(a), Isolation and characterization of resistant bacteria to heavy metals from mining areas of Zaire. Arch. Int. Physiol. Biochim. 96(2) B83; Diels et al., 1988(b), Detection of heterotrophic bacteria with plasmid-bound resistances to hea
REFERENCES:
Boylan et al., Lux C, D and E Genes of the Vibrio Fischeri Luminescence Operon Code for the Reductase, Transferase, and Synthetase Enzymes Involved in Aldehyde Biosynthesis, Chemical Abstracts, 111, Abs. No. 91344q, 189 (1989).
Fernando et al., "Cloning and Expression of an Avian Metallothionein-Encoding Gene", Gene, 81, 177-183 (1989).
Furukawa et al., "Molecular Relationship of Chromosomal Genes Encoding Biphenyl/Polychlorinated Biphenyl Catabolism: Some Soil Bacteria Possess a Highly Conserved bph Operon", Journal of Bacteriology, 171, 5467-5472 (1989).
Nies et al., Journal of Bacteriology, 169(10):4865-4868, 1987.
Shields et al., Journal of Bacteriology, 163(3):882-889, 1985.
Nies et al., Journal of Bacteriology, 171(2):896-900, 1989.
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D. Nies, et al. J. Bacteriol 174: 8102 (1992).
Corbisier Philippe
Diels Ludovicus
Mergeay Maximilien
Robinson Douglas W.
Vito
Wai Thanda
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