1887

Abstract

Previously, a gene encoding a predicted 67 kDa membrane-associated ATPase subunit of the Clp protease (ClpX) was identified in a porcine strain (95/1000) of the intestinal spirochaete In the current study, the distribution of this large gene was investigated in a collection of strains representing all seven spp. Using PCR with internal primers, an 878 bp portion of the gene was detected in 29 of 35 strains (83 %) of , 6 of 24 strains (25 %) of , 14 of 16 strains (88 %) of , 6 of 17 strains (35 %) of , 1 of 6 strains (17 %) of , 1 of 2 strains (50 %) of and not in the single strain of . The whole gene was sequenced from 20 spp. strains and compared with the gene from 95/1000 (GenBank accession no. AY466377). The genes had 99.3–99.7 % nucleotide sequence similarity and the predicted products had 99.7–100 % amino acid sequence similarity. The gene from WesB, a human strain of , was cloned and expressed as a histidine-tagged fusion protein in BL21. The purified protein was used to vaccinate mice and their sera were found to recognize the expected ∼67 kDa protein in whole-cell preparations of WesB. Sera from mice vaccinated with formalin-treated whole-cell proteins of WesB reacted with the recombinant protein. These results indicate that ClpX is both conserved and immunogenic and hence might be useful as a subunit vaccine component for spp. infections. Sera from humans with no known exposure to reacted with the recombinant ClpX protein, indicating that it is unlikely to be useful as a reagent for serological detection of spp. infections.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.47004-0
2007-07-01
2019-11-13
Loading full text...

Full text loading...

/deliver/fulltext/jmm/56/7/930.html?itemId=/content/journal/jmm/10.1099/jmm.0.47004-0&mimeType=html&fmt=ahah

References

  1. Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. ( 1997; ). Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402.[CrossRef]
    [Google Scholar]
  2. Bendtsen, J. D., Nielsen, H., Heijne, G. & Brunak, S. ( 2004; ). Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340, 783–795.[CrossRef]
    [Google Scholar]
  3. Brook, C. J., Clair, A. N., Mikosza, A. S. J., Riley, T. V. & Hampson, D. J. ( 2001; ). Carriage of intestinal spirochaetes by humans: epidemiological data from Western Australia. Epidemiol Infect 127, 369–374.
    [Google Scholar]
  4. Gardy, J. L., Laird, M. R., Chen, F., Rey, S., Walsh, C. J., Ester, M. & Brinkman, F. S. L. ( 2005; ). PSORTb v.2.0: expanded prediction of bacterial protein subcellular localization and insights gained from comparative proteome analysis. Bioinformatics 21, 617–623.[CrossRef]
    [Google Scholar]
  5. Gottesman, S., Clark, W. P., Lagard, V. C. & Maurizi, M. R. ( 1993; ). ClpX, an alternative subunit for the ATP-dependent Clp protease of Escherichia coli.. J Biol Chem 268, 22618–22626.
    [Google Scholar]
  6. Hampson, D. J. & Duhamel, G. E. ( 2006; ). Porcine colonic spirochetosis/intestinal spirochetosis. In Diseases of Swine, 9th edn, pp. 755–767. Edited by B. E. Straw, J. J. Zimmerman, S. D'Allaire & D. J. Taylor. Oxford: Blackwell Publishing.
  7. Hampson, D. J. & Stanton, T. B. (editors) ( 1997; ). Intestinal Spirochaetes in Domestic Animals and Humans. Wallingford, UK: CABI.
  8. Hampson, D. J., Robertson, I. D., La, T., Oxberry, S. L. & Pethick, D. W. ( 2000; ). Influences of diet and vaccination on colonisation of pigs by the intestinal spirochaete Brachyspira (Serpulina) pilosicoli. Vet Microbiol 73, 75–84.[CrossRef]
    [Google Scholar]
  9. Hampson, D. J., Fellström, C. & Thomson, J. R. ( 2006a; ). Swine dysentery. In Diseases of Swine, 9th edn, pp. 785–805. Edited by B. E. Straw, J. J. Zimmerman, S. D'Allaire & D. J. Taylor. Oxford: Blackwell Publishing.
  10. Hampson, D. J., Oxberry, S. L. & La, T. ( 2006b; ). Potential for zoonotic transmission of Brachyspira pilosicoli.. Emerg Infect Dis 12, 869–870.[CrossRef]
    [Google Scholar]
  11. Harland, W. A. & Lee, F. D. ( 1967; ). Intestinal spirochaetosis. Brit Med J 3, 718–719.[CrossRef]
    [Google Scholar]
  12. Hofmann, K. & Stoffel, W. ( 1993; ). Tmbase – a database of membrane spanning proteins segments. Biol Chem Hoppe Seyler 347, 166
    [Google Scholar]
  13. Kumar, S., Tamura, K. & Nei, M. ( 2004; ). mega3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5, 150–163.[CrossRef]
    [Google Scholar]
  14. Kunkle, R. A., Harris, D. L. & Kinyon, J. M. ( 1986; ). Autoclaved liquid medium for propagation of Treponema hyodysenteriae.. J Clin Microbiol 24, 669–671.
    [Google Scholar]
  15. Marchler-Bauer, A., Panchenko, A. R., Shoemaker, B. A., Thiessen, P. A., Geer, L. Y. & Bryant, S. H. ( 2002; ). CDD: a database of conserved domain alignments with links to domain three-dimensional structure. Nucleic Acids Res 30, 281–283.[CrossRef]
    [Google Scholar]
  16. Marchler-Bauer, A., Anderson, J. B., Cherukuri, P. F., DeWeese-Scott, C., Geer, L. Y., Gwadz, M., He, S., Hurwitz, D. I., Jackson, J. D. & other authors ( 2005; ). CDD: a Conserved Domain Database for protein classification. Nucleic Acids Res 33, D192–D196.[CrossRef]
    [Google Scholar]
  17. Mikosza, A. S. J. & Hampson, D. J. ( 2001; ). Human intestinal spirochetosis: Brachyspira aalborgi and/or Brachyspira pilosicoli?. Anim Health Res Rev 2, 101–110.
    [Google Scholar]
  18. Skinner, M. M. & Trempy, J. E. ( 2001; ). Expression of clpX, an ATPase subunit of the clp protease, is heat and cold shock inducible in Lactococcus lactis.. J Dairy Sci 84, 1783–1785.[CrossRef]
    [Google Scholar]
  19. Trott, D. J., Alt, D. P., Zuerner, R. L., Wannemuehler, M. J. & Stanton, T. B. ( 2001; ). The search for Brachyspira outer membrane proteins that interact with the host. Anim Health Res Rev 2, 19–30.
    [Google Scholar]
  20. Trott, D. J., Alt, D. P., Bulach, D. M., Zuerner, R. L. & Stanton, T. B. ( 2003; ). Identification of a membrane-bound clip protease (ClpX) homologue in Brachyspira pilosicoli. In Proceedings of the 2nd International Conference on Colonic Spirochaetal Infections in Animals and Humans, Eddlestone, Scotland. Abstract 59.
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.47004-0
Loading
/content/journal/jmm/10.1099/jmm.0.47004-0
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