Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Sep;177(17):4836–4840. doi: 10.1128/jb.177.17.4836-4840.1995

Isolation and sequencing of the replication region of Mycobacterium avium plasmid pLR7.

M L Beggs 1, J T Crawford 1, K D Eisenach 1
PMCID: PMC177255  PMID: 7665458

Abstract

The Mycobacterium avium plasmid pLR7 is representative of a group of small plasmids that are common in isolates from AIDS patients with disseminated M. avium infections. Determination of the functions of these and other plasmids has been hampered by the lack of methods for genetic manipulation of M. avium. In this study, the region of pLR7 capable of replication was identified and sequenced. Fragments of pLR7 were cloned into a pUC18 derivative carrying a kanamycin resistance marker and introduced into a plasmid-free M. avium strain by electroporation. The origin of replication was located on a 1.8-kb PvuII-to-SmaI fragment. An open reading frame encoding a putative Rep protein was identified. Two other open reading frames were identified in this region. A shuttle vector, pMB351, was constructed with the pLR7 origin of replication, pUC18, and the kanamycin resistance gene from Tn5. This vector was successfully transformed into M. avium, Mycobacterium tuberculosis, and Mycobacterium bovis.

Full Text

The Full Text of this article is available as a PDF (242.3 KB).

Selected References

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

  1. Bramhill D., Kornberg A. A model for initiation at origins of DNA replication. Cell. 1988 Sep 23;54(7):915–918. doi: 10.1016/0092-8674(88)90102-x. [DOI] [PubMed] [Google Scholar]
  2. Brennan R. G., Matthews B. W. The helix-turn-helix DNA binding motif. J Biol Chem. 1989 Feb 5;264(4):1903–1906. [PubMed] [Google Scholar]
  3. Crawford J. T., Bates J. H. Analysis of plasmids in Mycobacterium avium-intracellulare isolates from persons with acquired immunodeficiency syndrome. Am Rev Respir Dis. 1986 Oct;134(4):659–661. doi: 10.1164/arrd.1986.134.4.659. [DOI] [PubMed] [Google Scholar]
  4. Crawford J. T., Bates J. H. Restriction endonuclease mapping and cloning of Mycobacterium intracellulare plasmid pLR7. Gene. 1984 Mar;27(3):331–333. doi: 10.1016/0378-1119(84)90079-9. [DOI] [PubMed] [Google Scholar]
  5. Crawford J. T., Cave M. D., Bates J. H. Characterization of plasmids from strains of Mycobacterium avium-intracellulare. Rev Infect Dis. 1981 Sep-Oct;3(5):949–952. doi: 10.1093/clinids/3.5.949. [DOI] [PubMed] [Google Scholar]
  6. Hatfull G. F., Sarkis G. J. DNA sequence, structure and gene expression of mycobacteriophage L5: a phage system for mycobacterial genetics. Mol Microbiol. 1993 Feb;7(3):395–405. doi: 10.1111/j.1365-2958.1993.tb01131.x. [DOI] [PubMed] [Google Scholar]
  7. Hayashi S., Wu H. C. Lipoproteins in bacteria. J Bioenerg Biomembr. 1990 Jun;22(3):451–471. doi: 10.1007/BF00763177. [DOI] [PubMed] [Google Scholar]
  8. Hellyer T. J., Brown I. N., Dale J. W., Easmon C. S. Plasmid analysis of Mycobacterium avium-intracellulare (MAI) isolated in the United Kingdom from patients with and without AIDS. J Med Microbiol. 1991 Apr;34(4):225–231. doi: 10.1099/00222615-34-4-225. [DOI] [PubMed] [Google Scholar]
  9. Kües U., Stahl U. Replication of plasmids in gram-negative bacteria. Microbiol Rev. 1989 Dec;53(4):491–516. doi: 10.1128/mr.53.4.491-516.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Manen D., Caro L. The replication of plasmid pSC101. Mol Microbiol. 1991 Feb;5(2):233–237. doi: 10.1111/j.1365-2958.1991.tb02103.x. [DOI] [PubMed] [Google Scholar]
  11. Pabo C. O., Sauer R. T. Protein-DNA recognition. Annu Rev Biochem. 1984;53:293–321. doi: 10.1146/annurev.bi.53.070184.001453. [DOI] [PubMed] [Google Scholar]
  12. Qin M., Taniguchi H., Mizuguchi Y. Analysis of the replication region of a mycobacterial plasmid, pMSC262. J Bacteriol. 1994 Jan;176(2):419–425. doi: 10.1128/jb.176.2.419-425.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ranes M. G., Rauzier J., Lagranderie M., Gheorghiu M., Gicquel B. Functional analysis of pAL5000, a plasmid from Mycobacterium fortuitum: construction of a "mini" mycobacterium-Escherichia coli shuttle vector. J Bacteriol. 1990 May;172(5):2793–2797. doi: 10.1128/jb.172.5.2793-2797.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Scott J. R. Regulation of plasmid replication. Microbiol Rev. 1984 Mar;48(1):1–23. doi: 10.1016/b978-0-12-048850-6.50006-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Snapper S. B., Melton R. E., Mustafa S., Kieser T., Jacobs W. R., Jr Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis. Mol Microbiol. 1990 Nov;4(11):1911–1919. doi: 10.1111/j.1365-2958.1990.tb02040.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES