1887

Abstract

SUMMARY: Purified isoenzymes of --arabinofuranosidase (AF I, pI value 3·0) and polygalacturonase (PG III, pI 5·5; PGIV, pI 9·7) separated from culture filtrates of by isoelectric focusing and gel chromatography were used to produce antisera in sheep. The serum produced to AF I was specific as judged by immunodiffusion, immunoadsorbent column chromatography and binding of fluorescein-conjugated serum to the antigen separated by polyacrylamide gel electrophoresis. Fluorescein-labelled anti-AF I serum was bound to mycelium producing this isoenzyme in liquid culture, and binding was decreased when AF production was repressed with glucose; no labelling occurred on mycelium infecting apple fruit tissue.

Data for the anti-PG sera were less conclusive. The antigens gave precipitin lines only with homologous sera, but an anti-PG III immunoadsorbent column bound PG III as well as a smaller proportion of PG IV, while an anti-PG IV column bound neither antigen. Fluorescein isothiocyanate-labelled anti-PG III serum was bound to mycelium of grown in liquid culture and to mycelium-infected apple fruit tissue, but labelled anti-PG IV serum was not bound.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-83-1-135
1974-07-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/83/1/mic-83-1-135.html?itemId=/content/journal/micro/10.1099/00221287-83-1-135&mimeType=html&fmt=ahah

References

  1. Avrameas S., Ternynck T. 1969; The cross-linking of proteins with gluteraldehyde and its use for the preparation of immunoadsorbents. Immunochemistry 6:53–66
    [Google Scholar]
  2. Byrde R. J. W., Fielding A. H. 1968; Pectin methyl-trans-eliminase as the maceration factor in Sclerotinia fructigena and its significance in brown rot of apple. Journal of General Microbiology 52:287–297
    [Google Scholar]
  3. Byrde R. J. W., Fielding A. H., Archer S. A., Davies E. 1973; The role of extracellular enzymes in the rotting of fruit tissue by Sclerotinia fructigena . In Fungal Pathogenicity and the Plant#x2019;s Response pp 19–38 Byrde R. J. W., Cutting C. V. Edited by London and New York: Academic Press;
    [Google Scholar]
  4. Clark H. F., Shepard C. C. 1963; A dialysis technique for preparing fluorescent antibody. Virology 20:642–644
    [Google Scholar]
  5. Dingle J., Reid W. W., Solomons G. L. 1953; The enzymic degradation of pectin and other polysaccharides. II. Application of the ‘cup-plate’ assay to the estimation of enzymes. Journal of the Science of Food and Agriculture 4:149–155
    [Google Scholar]
  6. Fielding A. H., Hough L. 1971; The synthesis of 2-naphthyl-α-l-arabinofuranoside. Carbohydrate Research 20:416–417
    [Google Scholar]
  7. Goldman M. 1968 Fluorescent Antibody Methods New York and London: Academic Press;
    [Google Scholar]
  8. Handbook of Experimental Immunology 1967 Weir D. C. Edited by Oxford and Edinburgh: Blackwell Scientific Publications;
  9. Hislop E. C., Barnaby V. M., Shellis C., Laborda F. 1974; Localization of α-l-arabinofuranosidase and acid phosphatase in mycelium of Sclerotinia fructigena . Journal of General Microbiology 81:79–99
    [Google Scholar]
  10. Laborda F., Archer S. A., Fielding A. H., Byrde R. J. W. 1974; Studies on the α-l-arabinofuranosidase complex from Sclerotinia fructigena in relation to brown rot of apple. Journal of General Microbiology 81:151–163
    [Google Scholar]
  11. Laborda F., Fielding A. H., Byrde R. J. W. 1973; Extra- and intracellular α-l-arabinofuranosidase of Sclerotinia fructigena . Journal of General Microbiology 79:321–329
    [Google Scholar]
  12. Lowry O. H., Rosebrough N. J., Farr A. C., Randall R. J. 1951; Protein measurements with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  13. Miller L. 1959; The use of dinitrosalicylic acid reagent for the determination of reducing sugars. Analytical Chemistry 31:426–428
    [Google Scholar]
  14. Newby T. J., Bourne F. J., Chidlow J. W., Steel M. J. 1974; A simple method for the preparation of antiserum to bovine immunoglobulins using immunosorbents. Research in Veterinary Science 15:
    [Google Scholar]
  15. Ouchterlony Ö. 1958; Diffusion-in-gel methods for immunological analysis. Progress in Allergy 5:1–78
    [Google Scholar]
  16. Porath J., Axén R., Ernbach S. 1967; Chemical coupling of proteins to agarose. Nature; London: 2151491–1492
    [Google Scholar]
  17. Wrigley C. W. 1969; Analytical fractionation of proteins according to isoelectric point by gel electro- focusing. Shandon Instrument Applications29
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-83-1-135
Loading
/content/journal/micro/10.1099/00221287-83-1-135
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