1887

Abstract

Summary: Electroporation with shuttle plasmids carrying a kanamycin resistance gene as a selectable marker failed to generate transformants in two mycobacterial species currently being used in human vaccine trials ( and vaccae). In contrast, efficient transformation [10-10 transformants (μg DNA)] was obtained using novel vectors with selection based on expression of resistance to hygromycin. The hygromycin resistance vector was also found to be more efficient than kanamycin resistance vectors for transformation of and . The hygromycin resistance vector was used to overexpress superoxide dismutase of in in a form suitable for detailed structural analysis. The potential use of this approach for generation of novel recombinant mycobacterial vaccines is discussed.

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1994-01-01
2021-08-05
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References

  1. Aldovini A., Young R.A. Humoral and cell—mediated immune responses to live recombinant BCG—HIV vaccines. Nature (1991); 351:479–482
    [Google Scholar]
  2. Cooper J.B., Driessen H.P.C.,, Wood S.P., Zhang Y., Young J. Crystallization and preliminary X-ray analysis of the superoxide dismutase from Mycobacterium tuberculosis. J Mol Bio 1994 in press;
    [Google Scholar]
  3. Garbe T., Harris D., Vordermeier M., Lathigra R., Ivanyi J., Young D. Expression of the Mycobacterium tuberculosis 19—kilodalton antigen in Mycobacterium smegmatis: immunological analysis and evidence of glycosylation.. Infect Immun (1993); 61:260–267
    [Google Scholar]
  4. Labidi A., David H.L., Roulland—Dussoix D. Cloning and expression of mycobacterial plasmid DNA in Escherichia coli. FEMS Microbiol Lett (1985); 30:221–225
    [Google Scholar]
  5. Labidi A., Mardis E., Roe B.A., Wallace R.J. Cloning and DNA sequence of the Mycobacterium fortuitum plasmid pAL5000. Plasmid (1992); 27:130–140
    [Google Scholar]
  6. Laemrnli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (1970); 227:680–685
    [Google Scholar]
  7. Lee M.H., Pascopella L., Jacobs W.R., Hatfull G. Sitespecific integration of mycobacteriophage L5: integration—pro¬ficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis, and bacille Calmette—Guerin. Proc Natl Acad Sci USA (1991); 88:3111–3115
    [Google Scholar]
  8. Lydiate D.J., Ashby A.M., Henderson D.J., Kieser H.M., Hopwood D.A. Physical and genetic characterization of chromosomal copies of the Streptomyces coelicolor mini-circle. J Gen Microbiol (1989); 135:941–955
    [Google Scholar]
  9. Radford A.J., Hodgson A.L.M. Construction and characterization of a Mycobacterium-Escherichia coli shuttle vector. Plasmid (1991); 25:149–153
    [Google Scholar]
  10. Ranes M.G., Rauzier J., Lagranderie M., Georghiou M., Gicquel B. Functional analysis of pAL5000, a plasmid from Mycobacterium fortuitum: construction of a ‘ mini ’ Mycobacterium- Escherichia coli shuttle vector. J Bacteriol (1990); 172:2293–2297
    [Google Scholar]
  11. Sambrook J., Fritsch E.F., Maniatis T. Molecular Cloning, A Laboratory Manual. 1989 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  12. Snapper S.B., Lugosi L., Jekkel A., Melton R.E., Koeser T., Bloom B.R., Jacobs W.R. Lysogeny and transformation in mycobacteria: stable expression of foreign genes. Proc Natl Acad Sci USA (1988); 85:6987–6991
    [Google Scholar]
  13. Snapper S.B., Melton R.E., Mustafa S., Kieser T., Jacobs W.R. Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis. Mol Microbiol (1990); 4:1911–1919
    [Google Scholar]
  14. Stanford J.L., Rook G.A.W.,, Bahr G.M., Dowlati Y., Ganapati R., Ghazi Saidi K., Lucas S., Ramu G., Torres P., Ho Minh Ly, Anstey N. Mycobacterium vaccae in immunoprophylaxis and immunotherapy of leprosy and tuberculosis. Vaccine (1990); 8:525–530
    [Google Scholar]
  15. Stover K.C., De la Cruz, V F, Fuerst T.R., Burlein J.E., Benson L.A., Bennet L.T., Bansal G.P., Young J.F., Lee M.H., Hatfull G.F., Snapper S.B., Barletta R.G., Jacobs W.R., Bloom B.R. New use of BCG for recombinant vaccines. Nature (1991); 351:456–460
    [Google Scholar]
  16. Talwar G.P. Concluding comments on development of a vaccine against leprosy. Lepr India (1978); 50:492–497
    [Google Scholar]
  17. Talwar G.P., Zaheer S.A., Mukherjee R., Walia R., Misra R.S., Sharma A.K. Immunotherapeutic effects of a vaccine based on a saprophytic cultivable mycobacterium, Mycobacterium w, in multibacillary leprosy patients. Vaccine (1990); 8:121–129
    [Google Scholar]
  18. Towbin H., Staehelin T., Gordon G. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA (1979); 76:4350–4354
    [Google Scholar]
  19. Zaheer S.A., Mukherjee R., Ramkumar B., Misra R.S., Sharma A.K., Kar H.K., Kaur H., Nair S., Mukherjee A., Talwar G.P. Combined multidrug and Mycobacterium w vaccine therapy in patients with multibacillary leprosy. Infect Dis (1993); 167:401–410
    [Google Scholar]
  20. Zhang Y., Lathigra R., Garbe T., Catty D., Young D. Genetic analysis of superoxide dismutase, the 23 kilodalton antigen of Mycobacterium tuberculosis. Mol Microbiol (1991); 5:381–391
    [Google Scholar]
  21. Zhang Y., Heym B., Allen B., Young D., Cole S. The catalase-peroxidase gene and isoniazid resistance of Mycobacterium tuberculosis. Nature (1992); 358:591–593
    [Google Scholar]
  22. Zhang Y., Garbe T., Young D. Transformation with katG restores isoniazid-sensitivity in Mycobacterium tuberculosis isolates resistant to a range of drug concentrations. Mol Microbiol (1993); 8:521–524
    [Google Scholar]
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