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. 1987 Jan;169(1):394–402. doi: 10.1128/jb.169.1.394-402.1987

Molecular cloning and expression of the 3-chlorobenzoate-degrading genes from Pseudomonas sp. strain B13.

M P Weisshaar, F C Franklin, W Reineke
PMCID: PMC211780  PMID: 3025183

Abstract

The genes specifying the utilization of 3-chlorobenzoate by Pseudomonas sp. strain B13 WR1 have been cloned by using a broad-host-range cosmid cloning system. Analysis of the catabolic products of the enzymatic reactions encoded by two hybrid cosmids, pMW65 and pMW90, by thin-layer and high-performance liquid chromatography demonstrated that both encoded the genes for the complete catabolism of 3-chlorobenzoate. Physical analysis of one of the cosmid derivatives, pMW65, by restriction endonuclease mapping and subcloning demonstrated that the pathway genes are encoded on a fragment no larger than 11 kilobases.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bagdasarian M., Lurz R., Rückert B., Franklin F. C., Bagdasarian M. M., Frey J., Timmis K. N. Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene. 1981 Dec;16(1-3):237–247. doi: 10.1016/0378-1119(81)90080-9. [DOI] [PubMed] [Google Scholar]
  2. Bayley S. A., Duggleby C. J., Worsey M. J., Williams P. A., Hardy K. G., Broda P. Two modes of loss of the Tol function from Pseudomonas putida mt-2. Mol Gen Genet. 1977 Jul 20;154(2):203–204. doi: 10.1007/BF00330838. [DOI] [PubMed] [Google Scholar]
  3. Bolivar F., Backman K. Plasmids of Escherichia coli as cloning vectors. Methods Enzymol. 1979;68:245–267. doi: 10.1016/0076-6879(79)68018-7. [DOI] [PubMed] [Google Scholar]
  4. Boulnois G. J. Colicin Ib does not cause plasmid-promoted abortive phage infection of Escherichia coli K-12. Mol Gen Genet. 1981;182(3):508–510. doi: 10.1007/BF00293944. [DOI] [PubMed] [Google Scholar]
  5. Chatterjee D. K., Chakrabarty A. M. Genetic homology between independently isolated chlorobenzoate-degradative plasmids. J Bacteriol. 1983 Jan;153(1):532–534. doi: 10.1128/jb.153.1.532-534.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chatterjee D. K., Chakrabarty A. M. Restriction mapping of a chlorobenzoate degradative plasmid and molecular cloning of the degradative genes. Gene. 1984 Feb;27(2):173–181. doi: 10.1016/0378-1119(84)90138-0. [DOI] [PubMed] [Google Scholar]
  7. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cooper R. A., Skinner M. A. Catabolism of 3- and 4-hydroxyphenylacetate by the 3,4-dihydroxyphenylacetate pathway in Escherichia coli. J Bacteriol. 1980 Jul;143(1):302–306. doi: 10.1128/jb.143.1.302-306.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Danbara H., Timmis J. K., Lurz R., Timmis K. N. Plasmid replication functions: two distinct segments of plasmid R1, RepA and RepD, express incompatibility and are capable of autonomous replication. J Bacteriol. 1980 Dec;144(3):1126–1138. doi: 10.1128/jb.144.3.1126-1138.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Don R. H., Pemberton J. M. Properties of six pesticide degradation plasmids isolated from Alcaligenes paradoxus and Alcaligenes eutrophus. J Bacteriol. 1981 Feb;145(2):681–686. doi: 10.1128/jb.145.2.681-686.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dorn E., Hellwig M., Reineke W., Knackmuss H. J. Isolation and characterization of a 3-chlorobenzoate degrading pseudomonad. Arch Microbiol. 1974;99(1):61–70. doi: 10.1007/BF00696222. [DOI] [PubMed] [Google Scholar]
  12. Dorn E., Knackmuss H. J. Chemical structure and biodegradability of halogenated aromatic compounds. Substituent effects on 1,2-dioxygenation of catechol. Biochem J. 1978 Jul 15;174(1):85–94. doi: 10.1042/bj1740085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dorn E., Knackmuss H. J. Chemical structure and biodegradability of halogenated aromatic compounds. Two catechol 1,2-dioxygenases from a 3-chlorobenzoate-grown pseudomonad. Biochem J. 1978 Jul 15;174(1):73–84. doi: 10.1042/bj1740073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Evans W. C., Smith B. S., Fernley H. N., Davies J. I. Bacterial metabolism of 2,4-dichlorophenoxyacetate. Biochem J. 1971 May;122(4):543–551. doi: 10.1042/bj1220543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Franklin F. C., Bagdasarian M., Bagdasarian M. M., Timmis K. N. Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and cloning of genes for the entire regulated aromatic ring meta cleavage pathway. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7458–7462. doi: 10.1073/pnas.78.12.7458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Frey J., Bagdasarian M., Feiss D., Franklin F. C., Deshusses J. Stable cosmid vectors that enable the introduction of cloned fragments into a wide range of gram-negative bacteria. Gene. 1983 Oct;24(2-3):299–308. doi: 10.1016/0378-1119(83)90090-2. [DOI] [PubMed] [Google Scholar]
  17. Ghosal D., You I. S., Chatterjee D. K., Chakrabarty A. M. Genes specifying degradation of 3-chlorobenzoic acid in plasmids pAC27 and pJP4. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1638–1642. doi: 10.1073/pnas.82.6.1638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Grund A. D., Gunsalus I. C. Cloning of genes for naphthalene metabolism in Pseudomonas putida. J Bacteriol. 1983 Oct;156(1):89–94. doi: 10.1128/jb.156.1.89-94.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hansen J. B., Olsen R. H. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. doi: 10.1128/jb.135.1.227-238.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Inouye S., Nakazawa A., Nakazawa T. Nucleotide sequence of the promoter region of the xylDEGF operon on TOL plasmid of Pseudomonas putida. Gene. 1984 Sep;29(3):323–330. doi: 10.1016/0378-1119(84)90061-1. [DOI] [PubMed] [Google Scholar]
  21. Jeenes D. J., Williams P. A. Excision and integration of degradative pathway genes from TOL plasmid pWW0. J Bacteriol. 1982 Apr;150(1):188–194. doi: 10.1128/jb.150.1.188-194.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Knackmuss H. J., Hellwig M. Utilization and cooxidation of chlorinated phenols by Pseudomonas sp. B 13. Arch Microbiol. 1978 Apr 27;117(1):1–7. doi: 10.1007/BF00689343. [DOI] [PubMed] [Google Scholar]
  23. Lehrbach P. R., Jeenes D. J., Broda P. Characterization by molecular cloning of insertion mutants in TOL catabolic functions. Plasmid. 1983 Mar;9(2):112–125. doi: 10.1016/0147-619x(83)90014-8. [DOI] [PubMed] [Google Scholar]
  24. Mermod N., Lehrbach P. R., Reineke W., Timmis K. N. Transcription of the TOL plasmid toluate catabolic pathway operon of Pseudomonas putida is determined by a pair of co-ordinately and positively regulated overlapping promoters. EMBO J. 1984 Nov;3(11):2461–2466. doi: 10.1002/j.1460-2075.1984.tb02156.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reineke W., Knackmuss H. J. Construction of haloaromatics utilising bacteria. Nature. 1979 Feb 1;277(5695):385–386. doi: 10.1038/277385a0. [DOI] [PubMed] [Google Scholar]
  26. Reineke W., Knackmuss H. J. Microbial metabolism of haloaromatics: isolation and properties of a chlorobenzene-degrading bacterium. Appl Environ Microbiol. 1984 Feb;47(2):395–402. doi: 10.1128/aem.47.2.395-402.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schmidt E., Knackmuss H. J. Chemical structure and biodegradability of halogenated aromatic compounds. Conversion of chlorinated muconic acids into maleoylacetic acid. Biochem J. 1980 Oct 15;192(1):339–347. doi: 10.1042/bj1920339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schreiber A., Hellwig M., Dorn E., Reineke W., Knackmuss H. J. Critical Reactions in Fluorobenzoic Acid Degradation by Pseudomonas sp. B13. Appl Environ Microbiol. 1980 Jan;39(1):58–67. doi: 10.1128/aem.39.1.58-67.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Williams P. A., Murray K. Metabolism of benzoate and the methylbenzoates by Pseudomonas putida (arvilla) mt-2: evidence for the existence of a TOL plasmid. J Bacteriol. 1974 Oct;120(1):416–423. doi: 10.1128/jb.120.1.416-423.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

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