1887

Abstract

SUMMARY: A rapid centrifugation method for purifying foot-and-mouth disease virus (FMDV) combined an isodensity separation below a moving zone separation in the same tube. Two ml. of crude infectious fluid were introduced over a 3 ml. nonlinear CsCl gradient in a swinging bucket rotor. This preformed gradient provided a gradual increase from density 1.0 to 1.3 g./ml. and a step from 1.3 to 1.6 g./ml. After centrifugation for 4 hr. at 37,000 rev./min. (120,000 g) and 4°, a 1 mm. wide light scattering zone was observed with type A virus near the bottom, clearly separated below debris extending from density 1.3 to the meniscus. The narrow light scattering zone contained (40 ± 12%) of the FMDV infectivity and its CsCl isodensity was 1.43 ± 0.01 g./ml. Southern bean mosaic virus and bacteriophage øX174 behaved similarly and were useful as density markers. Virus suspensions concentrated an average of 8 fold retained (47 ± 16) % of their infectious units and were studied in the analytical ultracentrifuge directly. Dialysed concentrated virus revealed characteristic particles in an electron microscope. Exposure to concentrated impure CsCl decreases the stability of the infectivity to such an extent that in 40% CsCl its half-life is about 4 days.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-27-2-231
1962-02-01
2021-10-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/27/2/mic-27-2-231.html?itemId=/content/journal/micro/10.1099/00221287-27-2-231&mimeType=html&fmt=ahah

References

  1. Anderson N. G. 1955; Studies on isolated cell components. VIII. High resolution gradient differential centrifugation. Exp. Cell Res 9:446
    [Google Scholar]
  2. Bachrach H. L., Breese S. S. jun., Callis J. J., Hess W. R., Patty R. E. 1957; Inactivation of foot-and-mouth disease virus by pH and temperature changes and by formaldehyde. Proc. Soc. exp. Biol., N.Y 95:147
    [Google Scholar]
  3. Bachrach H. L., Callis J. J., Hess W. R., Patty R. E. 1957; A plaque assay for foot-and-mouth disease virus and kinetics of virus reproduction. Virology 4:224
    [Google Scholar]
  4. Bachrach H. L., Breese S. S. jun. 1958; Purification and electron microscopy of foot-and-mouth disease virus. Proc. Soc. exp. Biol., N.Y 97:659
    [Google Scholar]
  5. Bradish C. J., Henderson W. M., Kirkham J. B. 1960; Concentration and electron microscopy of the characteristic particle of foot-and-mouth disease. J. gen. Microbiol 22:379
    [Google Scholar]
  6. Brakke M. K. 1961; Density gradient centrifugation and its application to plant viruses. Advanc. Virus Res 7:193
    [Google Scholar]
  7. Breese S. S. jun., Trautman R., Bachrach H. L. 1960; Analysis by electron microscopy and infectivity of foot-and-mouth disease virus in moving boundary and zone ultracentrifugation. Arch. Biochem. Biophys 87:1
    [Google Scholar]
  8. DeDuve C., Berthet J., Beaufay H. 1959; Gradient centrifugation of cell particles: theory and applications. In Progress in Biophysics and Biophysical Chemistry Ed. Butler J. A. V., Katz B. London: Pergamon Press;
    [Google Scholar]
  9. Graves J. H., Poppensiek G. C. 1960; Determination of the optimum age range of mice for use in experimental studies with foot-and-mouth disease virus. Amer. J. Vet. Res 21:694
    [Google Scholar]
  10. Jacobsen C. F., Linderstrom-lang K. 1940; Method for rapid determination of specific gravity. Acta physiol, scand 1:149
    [Google Scholar]
  11. Lindgren F. T., Nichols A. V. 1960; Structure and function of human serum lipoproteins. In The Plasma Proteins vol. II Ed. Putnam F. W. New York: Academic Press;
    [Google Scholar]
  12. Matthews R. E. F. 1959; Turnip yellow mosaic virus nucleoprotein particles with differing biological and physical properties. Nature, Lond 184:530
    [Google Scholar]
  13. Matthews R. E. F. 1960; Properties of nucleoprotein fractions isolated from turnip yellow mosaic virus preparations. Virology 12:521
    [Google Scholar]
  14. Meselson M., Stahl F. W., Vinograd J. 1957; Equilibrium sedimentation of macromolecules in density gradients. Proc. nat. Acad. Sci., Wash 43:581
    [Google Scholar]
  15. Miller G. L., Gasek J. McG. 1960; Drift of drops in density gradient columns. Anal. Biochem 1:78
    [Google Scholar]
  16. Pyl G. 1953; Die Herstellung eines Konzentrat-Adsorbat-Impfstoffes gegen Maul- und-Klauenseuche aus gereinigtem Antigen. Arch. exp. Vet. Med. (Leipzig) 7:238
    [Google Scholar]
  17. Sinsheimer R. L. 1959; Purification and properties of bacteriophage øX-174. J. mol. Biol 1:37
    [Google Scholar]
  18. Skinner H. H., Henderson W. M., Brooksby J. B. 1952; Use of unweaned white mice in foot-and-mouth disease research. Nature, Lond 169:794
    [Google Scholar]
  19. Spinco Technical Reviews 1960 An introduction to density gradient centrifugation Beckman Instruments, Inc., Spinco Division; Palo Alto, Calif:
    [Google Scholar]
  20. Strohmaier K., Mussgay M. 1959; Bestimmung der Sedimentationskonstante eines infektiosen Prinzips mit Nucleinsaurecharakter aus dem Virus der Maul-und-Klauenseuche mit Hilfe der Gradienten-Zentrifugation. Z. Naturf 146:171
    [Google Scholar]
  21. Thompson W. R. 1947; Use of moving averages and interpolation to estimate median-effective dose. Bad. Rev 11:115
    [Google Scholar]
  22. Trautman R. 1956; Operating and comparating procedures facilitating schlieren pattern analysis in analytical ultracentrifugation. J. phys. Chem 60:1211
    [Google Scholar]
  23. Trautman R., Breese S. S. jun. 1959; Moving boundary theory applied to preparative ultracentrifugation. J. phys. Chem 63:1592
    [Google Scholar]
  24. Trautman R., Savan M., Breese S. S. jun. 1959; Partition by zone ultracentrifugation of the two complement-fixing particles in the foot-and-mouth disease virus system. J. Amer. chem. Soc 81:4040
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-27-2-231
Loading
/content/journal/micro/10.1099/00221287-27-2-231
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