1887

Abstract

Different parts of the plasmid pAL5000 necessary for plasmid replication and incompatibility were defined and studied. Two ORFs, named and were defined which are necessary for replication. A pAL5000 derivative deleted in these genes can be made to replicate by providing the gene products A 435 bp fragment was defined which was necessary for replication and which had an influence on copy number. This region (), which contains several repeated motifs, was also able to confer a degree of incompatibility when cloned into an otherwise unrelated mycobacterial replicon. pAL5000-derived plasmids carrying two copies of the region had a lower copy number and were less stable than the wild-type. These effects were only observed when the two regions were in the same orientation. Plasmids carrying only the region and no other parts of pAL5000 could be made to replicate if and were supplied from another plasmid. Based on these findings, systems for selectively curing cells of one plasmid of a pair were designed and shown to be functional in These have potential as a simple delivery system for achieving transposon mutagenesis or gene replacement in mycobacteria.

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1996-10-01
2024-12-02
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References

  1. Abeles A. L., Snyder K. M., Chattoraj D. K. 1984; P1 plasmid replication: replicon structure. J Mol Biol 173:307–324
    [Google Scholar]
  2. Beggs M. L., Crawford J. T., Eisenach K. D. 1995; Isolation and sequencing of the replication region of Mycobacterium avium plasmid pLR7. J Bacterial 177:4836–4840
    [Google Scholar]
  3. Berg D. E., Davies J., Allet B., Rochaix J. D. 1975; Transposition of R factor genes to bacteriophage lambda. Proc Natl Acad Sci USA 723628–3632
    [Google Scholar]
  4. Berg D. E., Jorgensen R., Davies J. 1978; Transposable kanamycin-neomycin resistance determinants. In Microbiology- 1978 pp. 13–15 Edited by Schlessinger D. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  5. Churchward G., Linder P., Caro L. 1983; The nucleotide sequence of replication and maintenance functions encoded by plasmid pSClOl. Nucleic Acids Res 11:5645–5659
    [Google Scholar]
  6. David M., Lubinsky-Mink S., Ben-Zvi A., Ulitzur S., Kuhn J., Suissa M. 1992; A stable Escherichia coli-Mycobacterium smegmatis plasmid shuttle vector containing the mycobacteriophage D29 origin. Plasmid 28:267–271
    [Google Scholar]
  7. Dellagostin O. A., Wall S., Norman E., O´Shaughnessy T., Dale J. W., McFadden J. 1993; Construction and use of integrative vectors to express foreign genes in mycobacteria. Mol Microbiol 10:983–993
    [Google Scholar]
  8. Gavigan J. A., Guilhot C., Gicquel B., Martin C. 1995; Use of conjugative and thermosensitive cloning vectors for transposon delivery to Mycobacterium smegmatis . FEMS Microbiol Lett 127:35–39
    [Google Scholar]
  9. Guilhot C., Gicquel B., Martin C. 1992; Temperature-sensitive mutants of the Mycobacterium plasmid pAL5000. FEMS Microbiol Lett 98:181–186
    [Google Scholar]
  10. Hanahan D., Jessee J., Bloom F. R. 1991; Plasmid trans formation of Escherichia coli and other bacteria. Methods Erzymol 204:63–114
    [Google Scholar]
  11. Hiraga S. I., Sugiyama T., Itoh T. 1994; Comparative analysis of the replicon regions of eleven ColE2-related plasmids. J Bacterial 176:7233–7243
    [Google Scholar]
  12. Kamio Y., Tabuchi A., Itoh Y., Katagiri H., Terawaki Y. 1984; Complete nucleotide sequence of mini-Rtsl and its copy mutant. J Bacterial 158:307–312
    [Google Scholar]
  13. Labidi A., David H. L., Roulland-Dussoix D. 1985; Cloning and expression of mycobacterial plasmid DNA in Escherichia coli . FEMS Microbiol Lett 30:221–225
    [Google Scholar]
  14. Labidi A., Mardis E., Roe B. A., Wallace R. J. Jr 1992; Cloning and DNA sequence of the Mycobacterium fortuitum var. fortuitum plasmid pAL5000. Plasmid 27:130–140
    [Google Scholar]
  15. Liu S. -K., Eisen J. A., Hanawalt P. C., Tessman I. 1993; recA mutations that reduce the constitutive coprotease activity of the RecA1202(Prtc) protein: possible involvement of interfilament association in proteolytic and recombination activities. J Bacterial 175:6518–6529
    [Google Scholar]
  16. Lydiate D. J., Ashby A. M., Henderson D. J., Kieser H. M., Hopwood D. A. 1989; Physical and genetic characterization of chromosomal copies of the Streptomyces coelicolor mini-circle. J Gen Microbiol 135:941–955
    [Google Scholar]
  17. McAdam R. A., Weisbrod T. R., Martin J., Scuderi J. D., Brown A. M., Cirillo J. D., Bloom B. R., Jacobs W. R. Jr 1995; In vivo growth characteristics of leucine and methionine auxotrophic mutants of Mycobacterium bovis BCG generated by transposon mutagenesis. Infect Immun 63:1004–1012
    [Google Scholar]
  18. Nomura N., Yamagishi H., Oka A. 1978; Isolation and characterization of transducing coliphage fd carrying a kanamycin transposon. Gene 3:39–51
    [Google Scholar]
  19. NordstrÖm K. 1993; Plasmid replication and maintenance. In Plasmids: a Practical Approach, 2nd edn. pp. 2–38 Edited by Hardy K. G. Oxford: Oxford University Press;
    [Google Scholar]
  20. NordstrÖm K., Austin S. J. 1989; Mechanisms that contribute to the stable segregation of plasmids. Annu Rev Genet 23:37–69
    [Google Scholar]
  21. Qin M., Taniguchi H., Mizuguchi Y. 1994; Analysis of the replication region of a mycobacterial plasmid, pMSC262. J Bacteriol 176:419–425
    [Google Scholar]
  22. Ranes M. G., Rauzier J., Lagranderie M., Gheorghiu M., Gicquel B. 1990; Functional analysis of pAL5000, a plasmid from Mycobacterium fortuitum: construction of a ‘mini’ mycobacterium- Escherichia coli shuttle vector. J Bacteriol 172:2793–2797
    [Google Scholar]
  23. Rauzier J., Moniz-Pereira J., Gicquel-Sanzey B. 1988; Complete nucleotide sequence of pAL5000 a plasmid from Mycobacterium fortuitum . Gene 71:315–321
    [Google Scholar]
  24. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Gloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 745463–5467
    [Google Scholar]
  26. Smith C. A., Thomas C. M. 1984; Nucleotide sequence of the trfA gene of broad host-range plasmid RK2. J Mol Biol 175:251–262
    [Google Scholar]
  27. Snapper S. B., Lugosi L., Jekkel A., Melton R. E., Kieser T., Bloom B. R., Jacobs W. R. Jr 1988; Lysogeny and transformation in mycobacteria: stable expression of foreign genes. Proc Natl Acad Sci USA 856987–6991
    [Google Scholar]
  28. Snapper S. B., Melton R. E., Mustafa S., Kieser T., Jacobs W. R. Jr 1990; Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis . Mol Microbiol 4:1911–1919
    [Google Scholar]
  29. Stover C. K., de la Cruz V. F., Fuerst T. R., Burlein J. E., Benson L. A., Bennett L. T., Bansal G. P., Young J. F., Lee M. H., Hatfull G. F., Snapper S. B., Barletta R. G., Jacobs W. R., Jacobs W. R. Jr Bloom B. R. 1991; New use of BCG for recombinant vaccines. Nature 351:456–460
    [Google Scholar]
  30. Tsutsui H., Fujiyama A., Murotsu T., Matsubara K. 1983; Role of nine repeating sequences of the mini-F genome for expression of F-specific incompatibility phenotype and copy number control. J Bacteriol 155:337–344
    [Google Scholar]
  31. West S. C. 1992; Enzymes and molecular mechanisms of genetic recombination. Annu Rev Biochem 61:603–640
    [Google Scholar]
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