1887

Abstract

Plasmid pEST4011 enables PaW85 to degrade 2,4-dichlorophenoxyacetic acid (2,4-D) and 3-chlorobenzoate (3-CBA). This new 2,4-D degradative plasmid has considerable homology with the regions of pJP4 containing the 2,4-D degradative genes . Restriction fragment HI-B of plasmid pEST4011, which has homology with this region, was cloned into the broad-host-range vector pKT240 and studied in PaW85. Restriction mapping, hybridization analysis and enzyme assays established the location of the genes for 2,4-D monooxygenase 2,4-dichlorophenol hydroxylase chlorocatechol 1,2-dioxygenase and the and regulatory genes on this fragment. Plasmid pEST4012 is a derivative of pEST4011 derived through the spontaneous deletion of a 42 kbp DNA fragment, which results in the loss of the 2,4-D and 3-CBA phenotype. We present here the physical maps of pEST4011 and pEST4012. In spite of the similarities in functions, the size (70 kbp), order of catabolic genes and restriction pattern of pEST4011 are clearly different from those of pJP4.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-12-3165
1993-12-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/12/mic-139-12-3165.html?itemId=/content/journal/micro/10.1099/00221287-139-12-3165&mimeType=html&fmt=ahah

References

  1. Amy P.S., Schulke J.W., Frazier L.M., Seidler R.J. 1985; Characterization of aquatic bacteria and cloning of genes specifying partial degradation of 2,4-dichlorophenoxyacetic acid. Appied and Environmental Microbiology 49:1237–1245
    [Google Scholar]
  2. Ausmees N.R., Heinaru A.L. 1990; New plasmids of herbicide 2,4-D degradation. Genetika 26:770–772 in Russian
    [Google Scholar]
  3. Bagdasarian M., Timmis K.N. 1982; Host vector systems for gene cloning in Pseudomonas. Current Topics of Microbiology and Immunology 96:46–67
    [Google Scholar]
  4. Bagdasarian M.M., Amann E., Lurz R., Rueckert B., Bagdasarian M. 1983; Activity of the hybrid trp.lac(tac) promotor of Escherichia coli in Pseudomonas putida. Construction of broad-host-range controlled-expression vectors. Gene 26:273–282
    [Google Scholar]
  5. Bauchop T., Elsden S.R. 1960; The growth of microorganisms in relation to their energy suppy. Journal of General Microbiology 23:469–495
    [Google Scholar]
  6. Bayley S.A., Duggleby C.J., Worsey M.J., Williams P.A., Hardy K.G., Broda P. 1977; Two modes of loss of the TOL function from Pseudomonas putida mt-2. Molecular and General Genetics 154:203–204
    [Google Scholar]
  7. Bradford M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  8. Chaudhry G.R., Huang G.H. 1988; Isolation and characterization of a new plasmid from Flavobacterium sp. which carries the genes for degradation of 2,4-dichlorophenoxyacetate. Journal of Bacteriology 170:3897–3902
    [Google Scholar]
  9. Connors M.A., Barnsley E.A. 1982; Naphthalene plasmids in Pseudomonas. Journal of Bacteriology 91:1140–1154
    [Google Scholar]
  10. Ditzelmüller G., Michael L., Streichsbier F. 1989; Isolation and characterization of a 2,4-dichlorophenoxyacetic acid-degrading soil bacterium. Appied Microbiology and Biotechnology 31:93–96
    [Google Scholar]
  11. Don R.H., Pemberton J.M. 1981; Properties of six pesticide degradation plasmids isolated from Alcaligenes eutrophus and Alcaligenes paradoxus. Journal of Bacteriology 145:681–686
    [Google Scholar]
  12. Don R.H., Pemberton J.M. 1985; Genetic and physical map of the 2,4-dichlorophenoxyacetic acid-degradative plasmid pJP4. Journal of Bacteriology 161:466–468
    [Google Scholar]
  13. Don R.H., Weightman A.J., Knackmuss H.J., Timmis K.N. 1985; Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134(pJP4). Journal of Bacteriology 161:85–90
    [Google Scholar]
  14. Downing R., Broda P.A. 1979; Cleavage map of the TOL plasmid of Pseudomonas putida mt-2. Molecular and General Genetics 111:189–191
    [Google Scholar]
  15. Ghosal D., You I.-S. 1988; Nucleotide homology and organization of chlorocatechol oxidation genes of plasmid pJP4 and pAC27. Molecular and General Genetics 211:113–120
    [Google Scholar]
  16. Ghosal D., You I.-S. 1989; Operon structure and nucleotide homology of the chlorocatechol oxidation genes of plasmids pJP4 and pAC27. Gene 83:225–232
    [Google Scholar]
  17. Ghosal D., You I.-S., Chatterjee D.K., Chakrabarty A.M. 1985; Plasmids in the degradation of chlorinated aromatic compounds. In Plasmids in Bacteria pp. 667–686 Helinski D.R., Cohen S.N., Clewell D.B., Jackson D.A., Hollaender A. Edited by New York: Plenum Press.;
    [Google Scholar]
  18. Gibson T.J. 1984 Studies on the Epstein-Barr virus genome. PhD thesis Cambridge University, Cambridge (UK):
    [Google Scholar]
  19. Hageman G.D. 1966; Synthesis of the enzymes of the mandelate pathway by Pseudomonas putida. I. Synthesis of the enzymes by the wild-type. Journal of Bacteriology 91:1140–1154
    [Google Scholar]
  20. Hanahan D. 1983; Studies on transformation of E. coli with plasmids. Journal of Molecular Biology 166:577–580
    [Google Scholar]
  21. Hansen J.E., Olsen R.H. 1978; Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pGMl and pGM5. Journal of Bacteriology 135:227–238
    [Google Scholar]
  22. Harker A.R., Olsen R.H., Seidler R.J. 1989; Phenoxyacetic acid degradation by the 2,4-dichlorophenoxyacetic acid (TFD) pathway of plasmids pJP4: mapping and characterization of the TFD regulatory gene tfdR. Journal of Bacteriology 171:314–320
    [Google Scholar]
  23. Kaphammer B., Olsen R.H. 1990; Cloning and characterization of tfdS, the repressor-activator gene of tfdB, from the 2,4-dichlorophenoxyacetic acid catabolic plasmid pJP4. Journal of Bacteriology 172:5856–5862
    [Google Scholar]
  24. Kaphammer B., Kukor J.J., Olsen R.H. 1990; Regulation of tfd CDEF by tfdR of 2,4-dichlorophenoxyacetic acid degradation plasmid pJP4. Journal of Bacteriology 172:2280–2286
    [Google Scholar]
  25. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning. A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  26. Pemberton J.M., Fisher P.R. 1977; 2,4-D plasmids and persistence. Nature: London; 268732–733
    [Google Scholar]
  27. Perkins E.J., Gordon M.P., Caceres D., Lurquin P.F. 1990; Organization and sequence analysis of the 2,4-dichlorophenol hydroxylase and dichlorocatechol oxidative operons of plasmid pJP4. Journal of Bacteriology 172:2351–2359
    [Google Scholar]
  28. Pierce G.E., Facklam T.J., Rice J.M. 1981; Isolation and characterization of plasmids from environmental strains of bacteria capable of degrading the herbicide 2,4-D. Developments in Industrial Microbiology 22:401–408
    [Google Scholar]
  29. Southern E.M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  30. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13 mpl8 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-12-3165
Loading
/content/journal/micro/10.1099/00221287-139-12-3165
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error