1887

Abstract

Scrutiny of sequence data from the genome sequencing project identified the presence of a gene encoding a 268-amino-acid polypeptide which is highly similar to a pore-forming haemolysin/cytotoxin virulence determinant, TlyA, from the swine pathogen Using degenerate oligonucleotide primers based on the TlyA sequences, the homologue was amplified and this product was used to obtain the clone and sequence a 2.5 kb fragment containing the whole gene. encodes a 267-amino-acid protein with a predicted molecular mass of 28 kDa. TlyA homologues were identified by PCR in and BCG, but appeared absent in and The gene appeared to be the first gene in an operon containing at least two other genes. Introduction of the gene into using a mycobacterial shuttle expression plasmid converted non-haemolytic cells into those exhibiting significant haemolytic activity. Similarly, inducible haemolytic activity was observed in sonicated bacteria when was expressed as a His-tagged fusion protein in mRNA was detected in both and BCG using RT-PCR, confirming that this gene is expressed in organisms cultured in

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1998-05-01
2021-08-04
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References

  1. Altschul, S. F., Gish, W., Miller, W., Myers, E. W., Lipman, D. J. (1990); Basic local alignment search tool.. Journal of Molecular Biology 215:(3)403–410 [View Article]
    [Google Scholar]
  2. Andrews, N. W., Portnoy, D. A. (1994); Cytolysins from intracellular pathogens.. Trends in Microbiology 2:(8)261–263 [View Article]
    [Google Scholar]
  3. Armstrong, J. A., Hart, P. D. (1971); Response of cultured macrophages to Mycobacterium tuberculosis, with observations on fusion of lysosomes with phagosomes.. Journal of Experimental Medicine 134:(3)713–740 [View Article]
    [Google Scholar]
  4. Bielecki, J., Youngman, P., Connelly, P., Portnoy, D. A. (1990); Bacillus subtilis expressing a haemolysin gene from Listeria monocytogenes can grow in mammalian cells.. Nature 345:(6271)175–176 [View Article]
    [Google Scholar]
  5. Clemens, D. L. (1996); Characterization of the Mycobacterium tuberculosis phagosome.. Trends in Microbiology 4:(3)113–118 [View Article]
    [Google Scholar]
  6. Deshpande, R. G., Khan, M. B.„ Bhat, D., A., Navalkar, R. G. (1997); Isolation of a contact-dependent haemolysin from Mycobacterium tuberculosis.. Journal of Medical Microbiology 46:(3)233–238 [View Article]
    [Google Scholar]
  7. Eiglmeier, K., Honore, N., Woods, S. A., Caudron, B., Cole, S. T. (1993); Use of an ordered cosmid library to deduce the genomic organization of Mycobacterium leprae.. Molecular Microbiology 7:(2)197–206 [View Article]
    [Google Scholar]
  8. Fischer, L. J., Quinn, F. D., White, E. H., King, C. H. (1996); Intracellular growth and cytotoxicity of Mycobacterium haemophilum in a human epithelial cell line (Hec-l-B).. Infection and Immunity 64:(1)269–276 [View Article]
    [Google Scholar]
  9. Flynn, J. L., Goldstein, M. M., Triebold, K. J., Koller, B., Bloom, B. R. (1992); Major histocompatibility complex class I-restricted T cells are required for resistance to Mycobacterium tuberculosis infection.. Proc Natl Acad Sci USA 89:(24)12013–12017 [View Article]
    [Google Scholar]
  10. ter Huurne, A. A., , Muir, S., van Houten, M., , van der Zeijst, B. A., , Gaastra, W. et al. (1994); Characterization of three putative Serpulina hyodysenteriae hemolysins.. Microb Pathog 16:(4)269–282 [View Article]
    [Google Scholar]
  11. Hyatt, D. R., Joens, L. A. (1997); Analysis of the lytic activity of the Serpulina hyodysenteriae hemolysin.. Infection and Immunity 65:(11)4877–4879 [View Article]
    [Google Scholar]
  12. Hyatt, D. R., ter Huurne, A. A., van der Zeijst, B. A., , Joens, L. A. (1994); Reduced virulence of Serpulina hyodysenteriae hemolysinnegative mutants in pigs and their potential to protect pigs against challenge with a virulent strain.. Infection and Immunity 62:(6)2244–2248 [View Article]
    [Google Scholar]
  13. Johansen, K. A., Gill, R. E., Vasin, M. L. (1996); Biochemical and molecular analysis of phospholipase C and phospholipase D activity in mycobacteria.. Infection and Immunity 64:(8)3259–3266 [View Article]
    [Google Scholar]
  14. King, C. H., Mundayoor, S., Crawford, J. T., Shinnick, T. M. (1993); Expression of contact-dependent cytolytic activity by Mycobacterium tuberculosis and isolation of the genomic locus that encodes the activity.. Infection and Immunity 61:(6)2708–2712 [View Article]
    [Google Scholar]
  15. Leao, S. C., Rocha, C. L., Murillo L.A., Parra, C. A., Patarroyo, M. E. (1995); A species-specific nucleotide sequence of Mycobacterium tuberculosis encodes a protein that exhibits hemolytic activity when expressed in Escherichia coli.. Infection and Immunity 63:(11)4301–4306 [View Article]
    [Google Scholar]
  16. McDonough, K. A., Kress, Y., Bloom, B. R. (1993); The interaction of Mycobacterium tuberculosis with macrophages: a study of phagolysosome fusion.. Infect Agents Dis 2:232–235
    [Google Scholar]
  17. Mangan, J. A., Sole, K. M., Mitchison, D. A., Butcher, P. D. (1997); An effective method of RNA extraction from bacteria refractory to disruption, including mycobacteria.. Nucleic Acids Research 25:(3)675–676 [View Article]
    [Google Scholar]
  18. Moreira, A. L., Wang, J., Tsenova Berkova, L., Hellmann, W., Freedman, V. H., Kaplan G. (1997); Sequestration of Mycobacterium tuberculosis in tight vacuoles in vivo in lung macrophages of mice infected by the respiratory route.. Infection and Immunity 65:(1)305–308 [View Article]
    [Google Scholar]
  19. Muir, S., Koopman, M. B., Libby, S. J., Joens, L. A., Heffron, F., Kusters J. G. (1992); Cloning and expression of a Serpula (Treponema) hyodysenteriae hemolysin gene.. Infection and Immunity 60:(2)529–535 [View Article]
    [Google Scholar]
  20. Porter, J. D. (1996); Mycobacteriosis and HIV infection: the new public health challenge.. J Antimicrob Chemother 37:(suppl B)113–120 [View Article]
    [Google Scholar]
  21. Rodrigues, L. C., Smith, P. G. (1990); Tuberculosis in developing countries and methods for its control.. Trans R Soc Trop Med Hyg 84:(5)739–744 [View Article]
    [Google Scholar]
  22. Sambrook, J., Fritsch, E. F., Maniatis, T. (1989) Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;
    [Google Scholar]
  23. Santos, A. R., De Miranda, A. B., Lima, L. M., Suffys, P. N., Degrave, W. M. (1992); Method for high yield preparation in large and small scale of nucleic acids from mycobacteria.. J Microbiol Methods 15:(2)83–94 [View Article]
    [Google Scholar]
  24. Smith, P. G., Moss, A. R. (1994) Epidemiology of tuberculosis. Edited by Bloom, B. R. Tuberculosis: Pathogenesis, Protection and Control Washington, DC:: American Society for Microbiology,;47–59
    [Google Scholar]
  25. 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.. Molecular Microbiology 4:(11)1911–1919 [View Article]
    [Google Scholar]
  26. Snider, D. E. Jr,, Raviglione M., Kochi A. (1994) Global burden of tuberculosis. Edited by Bloom, B. R. Tuberculosis: Pathogenesis, Protection and Control Washington, DC:: American Society for Microbiology,;3–11
    [Google Scholar]
  27. Stover, C. K., de la Cruz, V. F., , Fuerst, T. (1991); R. and 11 other authors, New use of BCG for recombinant vaccines.. Nature 351:(6326)456–460 [View Article]
    [Google Scholar]
  28. Thompson, J. D., Higgins, D. G., Gibson, T. J., Turner, J., Dockrell, H. M. (1994); clustal w : improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice, Stimulation of human peripheral blood mononuclear cells with live Mycobacterium bovis BCG activates cytolytic CD8 + T cells in vitro.. Nucleic Acids Research 22:(22)4673–4680 [View Article]
    [Google Scholar]
  29. Udou, T. (1994); Extracellular hemolytic activity in rapidly growing mycobacteria.. Canadian Journal of Microbiology 40:(4)318–321 [View Article]
    [Google Scholar]
  30. Wren, B. W., Colby, S. M., Cubberley, R. R., Pallen, M. J. (1992); Degenerate PCR primers for the amplification of fragments from genes encoding response regulators from a range of pathogenic bacteria.. Fems Microbiology Letters 78:(2–3)287–291 [View Article]
    [Google Scholar]
  31. Yang, J. W., Dhamija, S. S., Schweingruber, M. E. (1991); Characterisation of the specific p-nitrophenylphosphatase gene and protein of Schizosaccharomyces pombe.. European Journal of Biochemistry 198:(2)493–497 [View Article]
    [Google Scholar]
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