1887

Abstract

Summary: Mycobacterial expression signals were cloned using specially constructed gene fusion shuttle plasmid probes carrying a truncated (β-galactosidase) gene which lacked a promoter, a ribosome binding site, and an ATG start codon. Libraries of mycobacteriophage Bxb1, L1 and TM4 DNAs were constructed, and introduced by electroporation into and the ‘bacille Calmette-Guérin’ (BCG). Clones carrying mycobacterial expression sequences were detected by their blue colour or characteristic fluorescence when plated on media containing chromogenic or fluorogenic substrates. Varying degrees of β-galactosidase expression were observed, and one Bxb1 expression signal was identified where β-galactosidase expression is repressed in phage lysogens.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-138-1-23
1992-01-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/138/1/mic-138-1-23.html?itemId=/content/journal/micro/10.1099/00221287-138-1-23&mimeType=html&fmt=ahah

References

  1. ALDOVINI A., Young R. A. 1991; Humoral and cell-mediated immune responses to live recombinant BCG-HIV vaccines. Nature, London 351:479–482
    [Google Scholar]
  2. ARRAJ J. A., MARINUS M. G. 1983; Phenotypic reversal in dam mutants of Escherichia coli K-12 by a recombinant plasmid containing the dam+ gene. Journal of Bacteriology 153:562–565
    [Google Scholar]
  3. BARLETTA R. G., SNAPPER S. B., CIRILLO J. D., CONNELL N. D., KIM D. D., JACOBS W. R., JR & Bloom B. R. 1990; Recombinant BCG as a candidate oral vaccine. Research in Microbiology 141:931–939
    [Google Scholar]
  4. BOLIVAR F., RODRIGUEZ R. L., BETLACH M. C., BOYER H. W. 1977; Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9. Gene 2:75–93
    [Google Scholar]
  5. CENTERS FOR DISEASE CONTROL 1987; Tuberculosis final data - United States 1986. Morbidity and Mortality Weekly Report 36:817–820
    [Google Scholar]
  6. CLARK-CURTISS J. E., JACOBS W. R., DOCHERTY M. A., RITCHIE M. R., CURTISS R. III 1985; Molecular analysis of DNA and construction of genomic libraries of Mycobacterium leprae . Journal of Bacteriology 161:1093–1102
    [Google Scholar]
  7. DAGERT M., ERLICH S. D. 1979; Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 9:23–28
    [Google Scholar]
  8. DAVIS R. W., BOTSTEIN D., ROTH J. R. 1980 Advanced Bacterial Genetics Cold Spring Harbor; New York: Cold Spring Harbor Laboratory:
    [Google Scholar]
  9. FINE P. E. M. 1988; BCG vaccination against tuberculosis and leprosy. British Medical Bulletin 44:691–703
    [Google Scholar]
  10. GROSSMAN A. D., ERIKSON J. W., GROSS C. A. 1984; The htpR gene product of E. coli is a sigma factor for heat-shock promoters. Cell 38:383–390
    [Google Scholar]
  11. GUSSIN G. N., JOHNSON A. D., PABO C. O., SAUER R. T. 1983 Repressor and Cro protein: structure, function, and role in lysogenization. In Lambda II pp 93–121 Edited by Hendrix R. W., Roberts J. W., Stahl F. W., Weisberg R. A. Cold Spring Harbor; New York: Cold Spring Harbor Laboratory:
    [Google Scholar]
  12. HAWLEY D. K., McCLURE W. R. 1983; Compilation and analysis of Escherichia coli promoters DNA sequences. Nucleic Acids Research 11:2237–2255
    [Google Scholar]
  13. JACOBS W. R., DOCHERTY M. A., CURTISS R. III, CLARK-CURTISS J. E. 1986; Expression of Mycobacterium leprae genes from a Stereptococcus mutans promoter in Escherichia coli K-12. Proceedings of the National Academy of Sciences of the United States of America 83:1926–1930
    [Google Scholar]
  14. JACOBS W. R., JR TUCKMAN M., BLOOM B. R. 1987; Introduction of foreign DNA into mycobacteria using a shuttle phasmid. Nature, London 327: 532–535
    [Google Scholar]
  15. JACOBS W. R., JR GANJAM K. V., CIRILLO J. D., PASCOPELLA L., SNAPPER S. B., UDANI R. A., JONES W., BARLETTA R. G., BLOOM B. R. 1991; Genetic systems for the mycobacteria. Methods in Enzymology 204:537–555
    [Google Scholar]
  16. JOINT INTERNATIONAL UNION AGAINST TUBERCULOSIS & WORLD HEALTH ORGANIZATION STUDY GROUP 1982; Tuberculosis control. Tubercle 63:157–169
    [Google Scholar]
  17. KIESER T., MOSS M. T., DALE J. W., HOPWOOD D. A. 1986; Cloning and expression of Mycobacterium bovis BCG DNA in Streptomyces lividans . Journal of Bacteriology 168:72–80
    [Google Scholar]
  18. LABIDI A., DAUGUET C., GOH K. S., DAVID H. 1984; Plasmid profiles of Mycobacterium fortuitum complex isolates. Current Microbiology 11:235–240
    [Google Scholar]
  19. LEE M. H., PASCOPELLA L., JACOBS W. R., JR, & HATFULL G. F. 1991; Site-specific integration of mycobacteriophage L5 : Integration proficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis bacille Calmette-Guerin. Proceedings of the National Academy of Sciences of the United States of America 88:3111–3115
    [Google Scholar]
  20. LENNOX E. S. 1955; Transduction of linked genetic characters of the host by bacteriophage PI. Virology 1:190–206
    [Google Scholar]
  21. LUGOSI L., JACOBS W. R., JR, & BLOOM B. R. 1989; Genetic transformation of BCG. Tubercle 70:159–170
    [Google Scholar]
  22. MANIATIS T., FRITSCH E. F., SAMBROOK J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor; New York : Cold Spring Harbor Laboratory:
    [Google Scholar]
  23. MILLER J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor; New York: Cold Spring Harbor Laboratory:
    [Google Scholar]
  24. OKA A., SUGISAKI H., TANAKAMI M. 1981; Nucleotide sequence of the kanamycin-resistance transposon Tn903. Journal of Molecular Biology 147:217–226
    [Google Scholar]
  25. PTASHNE M., BACKMAN K., HUMAYAN Z., JEFFREY A., MAURER R., MEYER B., SAUER R. T. 1976; Autoregulation and function of a repressor in bacteriophage lambda. Science 194:156–161
    [Google Scholar]
  26. 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]
  27. RIGBY P. W. J., DIECKMANN M., RHODES C., BERG P. 1977; Labelling deoxyribonucleic acid to high specific activity in vitro by nick-translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  28. ROSENBERG M., COURT D. 1979; Regulatory sequences involved in the promotion and termination of transcription. Annual Review of Genetics 13:319–353
    [Google Scholar]
  29. SHINE J., DALGARNO L. 1974; The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA : complementarity to non-sense triplets and ribosome-binding sites. Proceedings of the National Academy of Sciences of the United States of America 71:1342–1346
    [Google Scholar]
  30. SILHAVY T. J., BERMAN M. L., ENQUIST L. W. 1984 Experiments with Gene Fusions Cold Spring Harbor; New York: Cold Spring Harbor Laboratory:
    [Google Scholar]
  31. SIRAKOVA T. D., BARDAROV S. S., KRIAKOV J. I., MARKOV K. I. 1989; Molecular cloning of mycobacterial promoters in Escherichia coli . FEMS Microbiology Letters 59:153–156
    [Google Scholar]
  32. 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. Proceedings of the National Academy of Sciences of the United States of America 85:6987–6991
    [Google Scholar]
  33. SNAPPER S. B., MELTON R. E., KIESER T., MUSTAFA S., JACOBS W. R. JR 1990; Isolation and characterization of high efficiency plasmid transformation mutants of Mycobacterium smegmatis . Molecular Microbiology 4:1911–1919
    [Google Scholar]
  34. 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. et al. 1991; Induction of antibody and T cell responses to foreign antigens by recombinant BCG vaccines. Nature, London 351:456–460
    [Google Scholar]
  35. THOLE J. R., DAUWERSE H. G., DAS P. K., GROOTHIUS D. G., SCHOULS L. M., VAN EMBDEN J. D. A. 1985; Cloning of Mycobacterium bovis BCG DNA and expression of antigens in Escherichia coli . Infection and Immunity 50:800–806
    [Google Scholar]
  36. TIMME T. L., BRENNAN P. J. 1984; Induction of bacteriophage from members of the Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium scrofulaceum serocomplex. Journal of General Microbiology 130:2059–2066
    [Google Scholar]
  37. WEINSTOCK G. M., BERMAN M. L., SILHAVY T. J. 1983 Chimeric genetics with β-galactosidase. In Gene Amplification and Analysis pp 27–64 Edited by Papas T. S., Rosenberg M., Chirikjian J. G. New York: Elsevier/North Holland Publishing Company;
    [Google Scholar]
  38. YOUNG R. A., BLOOM B. R., GROSSKINSKY C. M., IVANYI J., THOMAS D., DAVIS R. W. 1985a; Dissection of Mycobacterium tuberculosis antigens using recombinant DNA. Proceedings of the National Academy of Sciences of the United States of America 82:2583–2587
    [Google Scholar]
  39. YOUNG R. A., MEHRA V., SWEETSER D., BUCHANAN T., CLARK-CURTISS J., DAVIS R. W., BLOOM B. R. 1985b; Genes for the major protein antigens of the leprosy parasite Mycobacterium leprae . Nature, London 316:450–452
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-138-1-23
Loading
/content/journal/micro/10.1099/00221287-138-1-23
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error