1887

Abstract

Washed cells of serotype 4b, grown in broth culture at 20 °C and at 37 °C, were examined by electron microscopy for the presence of flagella. Many flagella were seen in cells grown at 20 °C, whereas at 37 °C very few were expressed. Flagella sheared from the cell surface were partially purified by differential centrifugation. Using SDS-PAGE and Western blotting two distinct protein bands were seen in this preparation, both with an apparent molecular mass of approximately 29 kDa. Further purification of these proteins was achieved by gel filtration and ion-exchange chromatography. Whole organisms grown at 20 °C and 37 °C were examined in Western blots using an affinity-purified polyclonal antibody, and a monoclonal antibody, both directed against 29 kDa putative flagellin. Bacteria grown at 20 °C expressed abundant flagellin, whereas only trace amounts could be detected in organisms grown at 37 °C. It is concluded that organisms grown at 20 °C both produce and assemble flagellin at the cell surface, and that flagellin production is a less marked feature of organisms grown at 37 °C.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-8-2171
1988-08-01
2022-01-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/8/mic-134-8-2171.html?itemId=/content/journal/micro/10.1099/00221287-134-8-2171&mimeType=html&fmt=ahah

References

  1. Abram D., Koffler H. 1964; In vitroformation of flagella-like filaments and other structures from flagellin. Journal of Molecular Biology 9:168–185
    [Google Scholar]
  2. Burnette W. N. 1981; ‘Western blotting’: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radiodinated protein A. Analytical Biochemistry 112:195–203
    [Google Scholar]
  3. Donker-Voet J. 1966; A serological study on some strains of Listeria monocytogenes . In Proceedings of the 3rd International Symposium on Listeriosis, Bilthoven, The Netherlands pp. 133–137
    [Google Scholar]
  4. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. 1977; Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature; London: 266550–552
    [Google Scholar]
  5. Herring A. J., Sharp J. M. 1984; Protein blotting: the basic method and its role in viral diagnosis. In Recent Advances in Virus Diagnosis pp. 115–125 Mcnulty M. S., Mcferran J. B. Edited by The Hague, Boston & London: Martinus Nijhoff;
    [Google Scholar]
  6. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  7. Leifson E., Palen M. I. 1955; Variations and spontaneous mutations in the genus Listeriain respect to flagellation and motility. Journal of Bacteriology 70:233–240
    [Google Scholar]
  8. Miles A. A., Misra S. S., Irwin J. O. 1938; The estimation of the bactericidal power of the blood. Journal of Hygiene 38:732–749
    [Google Scholar]
  9. Miller H. R. P., Ternynck T., Avrameas S. 1975; Synthesis of antibody and immunoglobulins without detectable antibody function in cells responding to horseradish peroxidase. Journal of Immunology 144:626–629
    [Google Scholar]
  10. Morrissey J. H. 1981; Silver stain for proteins on polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Analytical Biochemistry 117:307–310
    [Google Scholar]
  11. Nossal G. J. V., Ada G. L. 1971; Antigens and the afferent limb of the immune response. In Antigens, Lymphoid Cells and the Immune Response pp. 13–16 Dixon F. J., Kunkel M. G. Edited by New York & London: Academic Press;
    [Google Scholar]
  12. Paterson J. S. 1940; The antigenic structure of organisms of the genus Listerella . Journal of Pathology and Bacteriology 51:427–436
    [Google Scholar]
  13. Reading C. L. 1982; Theory and method of immunization in culture and monoclonal antibody production. Journal of Immunological Methods 53:261–291
    [Google Scholar]
  14. Richardson K., Parker C. D. 1985; Identification and occurrence of Vibrio choleraeflagellar core proteins in isolated outer membrane. Infection and Immunity 47:674–679
    [Google Scholar]
  15. Rose A. H. 1976 In An Introduction to Microbial Physiology, 3rd edn. pp. 16–18 London:: Butterworth.;
    [Google Scholar]
  16. Schmitt R., Raska I., Mayer F. 1974; Plain and complex flagella of Pseudomonas rhodos: analysis of the fine structure and composition. Journal of Bacteriology 117:844–857
    [Google Scholar]
  17. Seeliger H. P. R., Hohne K. 1979; Serotyping of Listeria monocytogenes and related species. Methods in Microbiology 13:31–49
    [Google Scholar]
  18. Whiteside T. M., Rhodes-Roberts M. E. 1985; Biochemical and serological properties of purified flagella and flagellins of some Pseudomonas spp. Journal of General Microbiology 131:873–883
    [Google Scholar]
  19. Wilson G. S., Miles A. A. 1964; Erysipelothrix and Listeria. In Topley and Wilson’s Principles of Bacteriology and Immunity, 5th edn. pp. 523–535 Wilson G. S., Miles A. A. Edited by London: Arnold;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-8-2171
Loading
/content/journal/micro/10.1099/00221287-134-8-2171
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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