Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 May;172(5):2793–2797. doi: 10.1128/jb.172.5.2793-2797.1990

Functional analysis of pAL5000, a plasmid from Mycobacterium fortuitum: construction of a "mini" mycobacterium-Escherichia coli shuttle vector.

M G Ranes 1, J Rauzier 1, M Lagranderie 1, M Gheorghiu 1, B Gicquel 1
PMCID: PMC208931  PMID: 2158981

Abstract

Functional domains of pAL5000 were determined by gene disruption and deletion analysis. Of the five plasmid open reading frames (ORFs), ORF1 to ORF5, and a putative origin of replication previously identified (J. Rauzier, J. Moniz-Pereira, and B. Gicquel-Sanzey, Gene 71:315-321), two of the ORFs (ORF3 and ORF4) were deemed dispensable for plasmid replication. A "mini" mycobacterium-Escherichia coli shuttle plasmid applicable for general recombinant DNA studies in mycobacteria was constructed by using the gene for Kanr (Tn903) as a selective marker. Heterologous expression of the gene for Kanr was confirmed by Western blotting (immunoblotting) analysis.

Full text

PDF
2794

Images in this article

Selected References

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

  1. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  2. Fine P. E. The BCG story: lessons from the past and implications for the future. Rev Infect Dis. 1989 Mar-Apr;11 (Suppl 2):S353–S359. doi: 10.1093/clinids/11.supplement_2.s353. [DOI] [PubMed] [Google Scholar]
  3. Gicquel-Sanzey B., Moniz-Pereira J., Gheorghiu M., Rauzier J. Structure of pAL5000, a plasmid from M. fortuitum and its utilization in transformation of mycobacteria. Acta Leprol. 1989;7 (Suppl 1):208–211. [PubMed] [Google Scholar]
  4. Jacobs W. R., Jr, Tuckman M., Bloom B. R. Introduction of foreign DNA into mycobacteria using a shuttle phasmid. Nature. 1987 Jun 11;327(6122):532–535. doi: 10.1038/327532a0. [DOI] [PubMed] [Google Scholar]
  5. Labidi A., David H. L., Roulland-Dussoix D. Restriction endonuclease mapping and cloning of Mycobacterium fortuitum var. fortuitum plasmid pAL5000. Ann Inst Pasteur Microbiol. 1985 Sep-Oct;136B(2):209–215. doi: 10.1016/s0769-2609(85)80045-4. [DOI] [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. McKenzie T., Hoshino T., Tanaka T., Sueoka N. The nucleotide sequence of pUB110: some salient features in relation to replication and its regulation. Plasmid. 1986 Mar;15(2):93–103. doi: 10.1016/0147-619x(86)90046-6. [DOI] [PubMed] [Google Scholar]
  8. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  9. Rauzier J., Moniz-Pereira J., Gicquel-Sanzey B. Complete nucleotide sequence of pAL5000, a plasmid from Mycobacterium fortuitum. Gene. 1988 Nov 30;71(2):315–321. doi: 10.1016/0378-1119(88)90048-0. [DOI] [PubMed] [Google Scholar]
  10. Snapper S. B., Lugosi L., Jekkel A., Melton R. E., Kieser T., Bloom B. R., Jacobs W. R., Jr Lysogeny and transformation in mycobacteria: stable expression of foreign genes. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6987–6991. doi: 10.1073/pnas.85.18.6987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES