1887

Abstract

SUMMARY

A method for the purification of enveloped infectious equine herpesvirus type 1 (EHV-1) is presented. Virus from cell culture fluids harvested at 48 h post infection was concentrated by sedimentation and partially purified by differential precipitation with ammonium sulphate. The final steps of purification consisted of two cycles of flotation of virus in pre-formed CsCl density gradients. Yields of infectious virus were about 30% (18 to 44%) of that present in starting material. As judged by electron microscopy, mixed radioisotope labelling, and absence of phosphohydrolase, virus preparations possessed a high degree of purity. Sedimentation of EHV-1 into CsCl density gradients resulted in low recovery of infectious virus. Flotation of virus in CsCl gradients, however, was not deleterious to infectivity of viral preparations.

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1976-04-01
2021-10-19
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References

  1. BOLLUM F. J. 1966 Procedures in Nucleic Acid Research section 3 296–306 Edited by Cantoni G. L., Davies D. R. New York and London: Harper and Row;
    [Google Scholar]
  2. BORGEN H. C., LUDWIG H. 1974; Equine herpesvirus 1: biological and biophysical comparison of two viruses from different clinical entities. Intervirology 4:189–198
    [Google Scholar]
  3. BRAY G. A. 1960; A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Analytical Biochemistry 1:279–285
    [Google Scholar]
  4. BURTON K. 1956; A study of the condition and mechanism of the diphenylamine reaction for the colori-metric determination of deoxyribonucleic acid. Biochemical Journal 62:315–323
    [Google Scholar]
  5. CRANE R. K., LIPMANN P. 1953; The effect of arsenate on aerobic phosphorylation. Journal of Biological Chemistry 201:235–243
    [Google Scholar]
  6. DISCHE Z. 1953; Qualitative and quantitative colorimetric determination of heptoses. Journal of Biological Chemistry 204:983–997
    [Google Scholar]
  7. GENTRY G. A., RANDALL C. C. 1973 The Herpesviruses chapter 3 45–92 Edited by Kaplan A. S. New York and London: Academic Press;
    [Google Scholar]
  8. GRAHAM B. J., LUDWIG H., BRONSON D. L., BENYESH-MELNICK M., BISWAL N. 1972; Physicochemical properties of the DNA of herpesviruses. Biochimica et Biophysica Acta 259:13–23
    [Google Scholar]
  9. IFFT J. B., VOET D. H., VINOGRAD J. 1961; The determination of density distributions and density gradients in binary solutions at equilibrium in the centrifuge. Journal of Physical Chemistry 65:1138–1145
    [Google Scholar]
  10. KEMP M. C, PERDUE M. L., RODGERS H. W., O’CALLAGHAN D. J., RANDALL C. C. 1974; Structural polypeptides of the hamster strain of equine herpes virus type 1: products associated with purification. Virology 61:361–375
    [Google Scholar]
  11. KLINGEBORN B., PERTOFT H. 1972; Equine abortion (Herpes) virus: purification and concentration of enveloped and de-enveloped virus and envelope material by density gradient centrifugation in colloidal silica. Virology 48:618–623
    [Google Scholar]
  12. LAWRENCE W. C., GINSBERG H. S. 1967; Intracellular uncoating of type 5 adenovirus deoxyribonucleic acid. Journal of Virology 1:851–867
    [Google Scholar]
  13. LOWRY O. H., ROSENBROUGH N. J., FARR A. L., RANDALL R. J. 1951; Protein measurements with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  14. MCCOMBS R. M. 1969; Concentration and purification of herpes viruses (simplex, cytomegalo and EB) in a zonal ultracentrifuge. Applied Microbiology 17:636–638
    [Google Scholar]
  15. PERDUE M. L., KEMP M. C., RANDALL C. C., O’CALLAGHAN D. J. 1974; Studies of the molecular anatomy of the L-M cell strain of equine herpes virus type 1: proteins of the nucleocapsid and intact virion. Virology 59:201–216
    [Google Scholar]
  16. PERTOFT S. 1970; Isopycnic zonal centrifugation in colloidal silica: concentration and purification of a herpesvirus (iBRV). Analytical Biochemistry 38:506–516
    [Google Scholar]
  17. RANDALL C. C., LAWSON L. A. 1962; Adaptation of equine abortion virus to Earle’s L cells in serum free medium. Proceedings of the Society for Experimental Biology and Medicine no:487–489
    [Google Scholar]
  18. RANDALL C. C, ROGERS H. W., DOWNER D. N., GENTRY G. A. 1972; Protein kinase activity in equine herpesvirus. Journal of Virology 9:216–222
    [Google Scholar]
  19. ROBINSON D. J., WATSON D. H. 1971; Structural proteins of herpes simplex virus. Journal of General Virology 10:163–171
    [Google Scholar]
  20. SHATKIN A. J. 1969 Fundamental Techniques in Virology chapter 23 238–241 Edited by Habel K., Salzman N. New York and London: Academic Press;
    [Google Scholar]
  21. SPEAR P. G., ROIZMAN B. 1972; Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpes virion. Journal of General Virology 22:297–302
    [Google Scholar]
  22. SPRING S. B., ROIZMAN B. 1967; Herpes simplex virus products in productive and abortive infection. I. Stabilization with formaldehyde and preliminary analyses by isopycnic centrifugation in CsCl. Journal of Virology I:294–301
    [Google Scholar]
  23. STECK T. L., STRAUS J. H., WALLACH D. F. H. 1970; A model for the behavior of vesicles in density gradients: implications for fractionation. Biochimica et Biophysica Acta 203:385–393
    [Google Scholar]
  24. VAHLNE A. G., BLOMBERG J. 1974; Purification of herpes simplex virus. Journal of General Virology 22:297–302
    [Google Scholar]
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