1887

Abstract

The dichloromethane (DCM)-utilizing facultative methylotroph sp. DM4 was shown to contain three plasmids with approximate sizes of 120 kb, 40 kb and 8 kb. Curing experiments suggested that the DCM-utilization character was correlated with the possession of an intact 120 kb plasmid. The DCM-utilization genes were cloned on the broad-host-range vector pVK100. Plasmid pME1510, a recombinant plasmid carrying a 21 kb dIII fragment complemented DCM-utilization-negative derivatives of sp. DM4 and conferred the DCM-utilization-positive phenotype to a number of Gram-negative methylotrophic bacteria. In Southern hybridization experiments with pME1510 as a probe, chromosomal DNA from sp. DM4 gave definite signals while purified plasmid DNA did not. Plasmid pME1510 did not hybridize with total DNA from a cured DCM-non-utilizing derivative of sp. DM4. It is concluded that the DCM-utilization genes are located on the chromosome or on a megaplasmid. Curing procedures thus led to the formation of a chromosomal or megaplasmid deletion larger than 21 kb and covering the DCM-utilization genes or to the loss of an undetected megaplasmid.

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1988-04-01
2024-04-27
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References

  1. Boyer H. W., Roulland-Russoix D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli. Journal of Molecular Biology 41:459–472
    [Google Scholar]
  2. Bradford M. 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]
  3. Brunner W., Staub D., Leisinger T. 1980; Bacterial degradation of dichloromethane. Applied and Environmental Microbiology 40:950–958
    [Google Scholar]
  4. Chesney R. H., Scott J. R., Vapnek D. 1979; Integration of the plasmid prophages P1 and P7 into the chromosome of Escherichia coli. Journal of Molecular Biology 130:161–173
    [Google Scholar]
  5. Figurski D. H., Helinski D. R. 1979; Replication of an orgin containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proceedings of the National Academy of Sciences of the United States of America 76:1648–1652
    [Google Scholar]
  6. GÄlli R., Leisinger T. 1985; Specialized bacterial strains for the removal of dichloromethane from industrial waste. Conservation and Recycling 8:91–100
    [Google Scholar]
  7. Green P. N., Bousfield I. J. 1982; A taxonomic study of some Gram-negative facultativelymethylo- trophic bacteria. Journal of General Microbiology 128:623–638
    [Google Scholar]
  8. Green P. N., Bousfield I. J. 1983; Emendation of MethylobacteriumPatt, Cole, and Hanson 1976; Methylobacterium rhodinum (Heumann 1962) comb, nov.corrig.;Methylobacterium radiotolerans (Ito and Iizuka 1971) comb. nov.corrig.; and Methylobacterium mesophilicum (Austin and Goodfellow 1979) comb. nov. International Journal of Systematic Bacteriology 33:875–877
    [Google Scholar]
  9. Haas D. 1983; Genetic aspects of biodegradation by pseudomonads. Experientia 38:1199–1213
    [Google Scholar]
  10. Haas D., Holloway B. W. 1978; Chromosome mobilization by the R plasmid R68.45: a tool in Pseudomonas genetics. Molecular and General Genetics 158:223–237
    [Google Scholar]
  11. Hohn B. 1973; In vitro packaging of λ and cosmid DNA. Methods in Enzymology 68:299–309
    [Google Scholar]
  12. Itoh Y., Watson J. M., Haas D., Leisinger T. 1984; Genetic and molecular characterization of the Pseudomonas plasmid pVSl. Plasmid 11:206–220
    [Google Scholar]
  13. Kado C. J., Liu S. T. 1981; Rapid procedure for detection and isolation of large and small plasmids. Journal of Bacteriology 145:1365–1373
    [Google Scholar]
  14. Kawasaki H., Tone N., Tonomura K. 1981; Plasmid-determined dehalogenation of haloacetates in Moraxella species. Agricultural and Biological Chemistry 45:29–34
    [Google Scholar]
  15. Knauf V. C., Nester E. W. 1982; Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid 8:45–54
    [Google Scholar]
  16. Kohler-Staub D., Leisinger T. 1985; Dichloromethane dehalogenase of Hyphomicrobium sp. strain DM2. Journal of Bacteriology 162:676–681
    [Google Scholar]
  17. Kohler-Staub D., Hartmans S., GÄlli R., Leisinger T. 1986; Evidence for identical dichloromethane dehalogenases in different methylo- trophic bacteria. Journal of General Microbiology 132:2837–2843
    [Google Scholar]
  18. Konheiser U., Bollschweiler C., Klein A. 1984; Physical mapping of genes coding for two subunits of methyl-CoMreductase component C of Methanococcus voltae. Molecular and General Genetics 198:146–152
    [Google Scholar]
  19. Lapat-Polasko L. T., Mccarty P. L., Zehnder A. J. B. 1984; Secondary substrate utilization of methylene chloride by an isolated strain of Pseudomonas sp. Applied and Environmental Microbiology 47:825–830
    [Google Scholar]
  20. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;
    [Google Scholar]
  21. Peel D., Quayle J. R. 1961; Microbial growth on Cl compounds. 1. Isolation and characterization of Pseudomonas AMI. Biochemical Journal 81:465–469
    [Google Scholar]
  22. Rigby P. W. J., Dieckman M., Rhodes C., Berg P. 1977; Labeling deoxyribonucleic acid to high specific activity in vitro by nick-translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  23. Schleif R. F., Wensink P. C. 1981 Practical Methods in Molecular Biology p. 201. New York:: Springer Verlag;
    [Google Scholar]
  24. Slater J. H., Weightman A. J., Hall B. G. 1985; Dehalogenase genes of Pseudomonas putidaPP3 on chromosomally located transposable elements. Molecular and Biological Evolution 2:557–567
    [Google Scholar]
  25. Smith G. E., Summers M. D. 1980; The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Analytical Biochemistry 109:123–129
    [Google Scholar]
  26. Stucki G., GÄlli R., Ebersold H.-R., Leisinger T. 1981; Dehalogenation of dichloromethane by cell extracts of Hyphomicrobium DM2. Archives of Microbiology 130:366–371
    [Google Scholar]
  27. Tatra P. K., Goodwin P. M. 1983; R-Plasmid- mediated chromosome mobilization in the facultative methylotroph Pseudomonas AMI. Journal of General Microbiology 129:2629–2632
    [Google Scholar]
  28. Wahl G. M., Stern M., Stark G. R. 1979; Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextransulfate. Proceedings of the National Academy of Sciences of the United States of America 76:3683–3687
    [Google Scholar]
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