1887

Abstract

pWW53-4 is a cointegrate between RP4 and the catabolic plasmid pWW53 from MT53, which contains 36 kbp of pWW53 DNA inserted close to the gene of RP4; it encodes the ability to grow on toluene and the xylenes, characteristic of pWW53, as well as resistance to tetracycline, kanamycin and carbenicillin, characteristic of RP4. A physical map of the 36 kbp insert of pWW53 DNA for 11 restriction enzymes is presented, showing that the relative positions of the two operons are different from those on the archetypal TOL plasmid pWW0. The location of the genes for 4-oxalocrotonate decarboxylase () and 4-oxalocrotonate tautomerase () were shown by subcloning and enzyme assay to lie at the distal end of the pathway operon. Although 2-oxopent-4-enoate hydratase () and 4-hydroxy-2-oxovalerate aldolase () could be detected on a large cloned III fragment, they could not be accurately located on smaller subcloned DNA, but the only credible position for them is between and . The gene order in the pathway operon is therefore (). The regulatory genes and were located close to and downstream of the pathway operon, and the restriction map of the DNA in this region, as has previously been shown for the two operons carrying the structural genes, shows similarities with the corresponding region on pWW0. Evidence is also presented for the existence of two promoters, termed P3 and P4, internal to the pathway operon which support low constitutive expression of the structural genes downstream in hosts but not in .

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1987-05-01
2022-09-27
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References

  1. Bagdasarian M., Lurz R., Rueckert B., Franklin F. C. H., Bagdasarian M. M., Frey J., Timmis K. N. 1981; Specific purpose cloning vectors. II. Broad host range, high copy number RSFlOlO-derived vectors and a host:vector system for gene cloning. Gene 16:237–247
    [Google Scholar]
  2. Bayly R. C., Dagley S., Gibson D. T. 1966; The metabolism of cresols by species of Pseudomonas. Biochemical Journal 101:293–301
    [Google Scholar]
  3. Cane P. A., Williams P. A. 1986; A restriction map of naphthalene catabolic plasmid pWW60-l and the location of some of its catabolic genes. Journal of General Microbiology 132:2919–2929
    [Google Scholar]
  4. Collinsworth W. L., Chapman P. J., Dagley S. 1973; Stereospecific enzymes in the degradation of aromatic compounds by Pseudomonas putida. Journal of Bacteriology 113:922–931
    [Google Scholar]
  5. Franklin F. C. H., Lehrbach P. R., Lurz R., Rueckert B., Bagdasarian M., Timmis K. N. 1983; Localisation and functional analysis of transposon mutations in regulatory genes of the TOL catabolic pathway. Journal of Bacteriology 154:676–685
    [Google Scholar]
  6. Grinter N. J. 1983; A broad host range cloning vector transposable to various replicons. Gene 21:133–143
    [Google Scholar]
  7. Harayama S., Lehrbach P. R., Timmis K. N. 1984; Transposon mutagenesis ofmeta-cleavage pathway operon genes of the TOL plasmid of Pseudomonas putida mt-2. Journal of Bacteriology 160:251–255
    [Google Scholar]
  8. Holmes D. S., Quigley M. 1981; A rapid boiling method for preparation of bacterial plasmids. Analytical Biochemistry 114:193–197
    [Google Scholar]
  9. Inouye S., Nakazawa A., Nakazawa T. 1983; Molecular cloning of regulatory gene xylR and operator promoter regions of xylABC and xylDEGF operons of the TOL plasmid. Journal of Bacteriology 155:1191–1199
    [Google Scholar]
  10. Keil H., Lebens M. R., Williams P. A. 1985a; TOL plasmid pW W15 contains two nonhomologous, independently regulated catechol 2,3-oxygenase genes. Journal of Bacteriology 163:248–255
    [Google Scholar]
  11. Keil H., Keil S., Pickup R. W., Williams P. A. 1985b; Evolutionary conservation of genes coding for meta pathway enzymes within TOL plasmids pWWO and pWW53. Journal of Bacteriology 164:887–895
    [Google Scholar]
  12. Keil H., Saint C. M., Williams P. A. 1987; Gene organization of first catabolic operon of TOL plasmid pWW53: production of indigo by xylA gene product. Journal of Bacteriology 169:764–770
    [Google Scholar]
  13. Kunz D. A., Chapman P. J. 1981; Isolation and characterization of spontaneously occurring TOL plasmid mutants of Pseudomonas putida HS1. Journal of Bacteriology 146:952–964
    [Google Scholar]
  14. Lanka E., Lurz E., Fuerste J.-P. 1983; Molecular cloning and mapping of SphI restriction fragments of plasmid RP4. Plasmid 10:303–307
    [Google Scholar]
  15. Lebens M. R., Williams P. A. 1985; Complemen-tation of deletion and insertion mutants of TOL plasmid pWWO: regulatory implications and loca-tion of xylC gene. Journal of General Microbiology 131:3261–3269
    [Google Scholar]
  16. Maniatis T., Fritsch E. F., Sambrook J. (eds) 1982 Molecular Cloning: a Laboratory Manual pp. 98–148 Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  17. Messing J. 1983; New Ml3 vectors for cloning. Methods in Enzymology 101:20–78
    [Google Scholar]
  18. Nakazawa T., Hayashi E., Yokota T., Ebina Y., Nakazawa A. 1978; Isolation of TOL and RP4 recombinants by integrative suppression. Journal of Bacteriology 134:270–277
    [Google Scholar]
  19. Prentki P., Kirsch A. M. 1982; A modified pBR322 vector with improved properties for the cloning, recovery and sequencing of blunt-ended DNA fragments. Gene 17:189–196
    [Google Scholar]
  20. Reiner A. M. 1972; Metabolism of aromatic compounds in bacteria. Purification and properties of the catechol-forming enzyme, 3,5-cyclohexadiene-1,2-diol-1-carboxylic acid (NAD+) oxidoreductase (decarboxylating). Journal of Biological Chemistry 247:4960–4965
    [Google Scholar]
  21. Reiner A. M., Hegeman G. D. 1971; Metabolism of benzoic acid by bacteria. Accumulation of (—)-3,5-cyclohexadiene-l,2-diol-1-carboxylic acid by a mutant strain of Alcaligenes eutrophus. Biochemistry 10:2530–2536
    [Google Scholar]
  22. Sala-Trepat J. M., Evans W. C. 1971; The meta cleavage of catechol by Azotobacter species: 4-oxalocrotonate pathway. European Journal of Biochemistry 20:400–413
    [Google Scholar]
  23. Spooner R. A., Lindsay K., Franklin F. C. H. 1986; Genetic, functional and sequence analysis of the xylR and xylS regulatory genes of the TOL plasmid pWWO. Journal of General Microbiology 132:1347–1358
    [Google Scholar]
  24. Williams P. A., Worsey M. J. 1976; Ubiquity of plasmids in coding for toluene and xylene metabolism in soil bacteria: evidence for the existence of new TOL plasmids. Journal of Bacteriology 125:818–828
    [Google Scholar]
  25. Worsey M. J., Williams P. A. 1975; Metabolism of toluene and xylenes by Pseudomonas putida (arvilla) mt-2: evidence for a new function of the TOL plasmid. Journal of Bacteriology 124:7–13
    [Google Scholar]
  26. Worsey M. J., Franklin F. C. H., Williams P. A. 1978; Regulation of the degradative pathway enzymes coded for by the TOL plasmid (pWWO) from Pseudomonas putida mt-2. Journal of Bacteriology 134:757–764
    [Google Scholar]
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