Summary
Conditions for growth, concentration, and purification of Herpesvirus saimiri were determined. Optimal yields of infectious Herpesvirus saimiri (HVS) were obtained from infected owl monkey kidney (OMK) cells grown at 32.5° C in medium containing 10 per cent fetal calf serum. Forty-five percent of the initial infectious HVS was recovered after an 18-fold concentration using 8 per cent polyethylene glycol 6000 in the presence of 0.5M NaCl. Polyethylene glycol concentrated HVS was purified in an isopycnic-linear Renografin gradient (1.0–1.3 g/cm3). Ninety-six percent of the infectivity was recovered in a single 1.16 g/cm3 density region. DNA extracted from purified HVS was resolved into two distinct density classes by CsCl equilibrium centrifugation (1.727 and 1.709 g/cm3). DNase treated HVS virions yield four DNA species with densities of 1.727, 1.718, 1.712, and 1.706 g/cm3 in CsCl centrifugation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adams, A.: Concentration of Epstein-Barr virus from cell culture fluids with polyethylene glycol. J. gen. Virol.20, 391–394 (1973).
Ablashi, D. V., Armstrong, G. R., Blackham, E. A.: Certain characteristics of Herpesvirus saimiri cultured in subhuman primate culture cells. Amer. J. vet. Res.33, 1689–1694 (1972).
Deinhardt, F.: In:Kaplan, A. S. (ed.), The Herpesviruses, 595–625. New York: Academic Press, 1973.
Fleckenstein, B., Bornkamm, G., Ludwig, H.: Repetitive sequences in complete and defective genomes ofHerpesvirus saimiri. J. Virol.15, 398–406 (1975).
Fleckenstein, B., Wolf, H.: Purification and properties ofHerpesvirus saimiri DNA. Virology58, 55–64 (1974).
Gentry, G. A., Bandall, C. C.: In:Kaplan, A. S. (ed.), The Herpesviruses, 45 to 92. New York: Academic Press, 1973.
Goodheart, C. R.: Herpesvirus and Cancer. J. Amer. Med. Ass.211, 91–96 (1970).
Helling, R. B., Goodman, H. M., Boyer, H. W.: Analysis of endonuclease R Eco RI fragments of DNA from lambdoid bacteriophages and other viruses by agarose-gel electrophoresis. J. Virol.14, 1235–1244 (1974).
Laufs, R., Fleckenstein, B.: Purification of Herpesvirus saimiri DNA. Bibl. Haematol.39, 457–461 (1973).
McSharry, J., Benzinger, R.: Concentration and purification of vesicular stomatitis virus by polyethylene glycol “precipitation”. Virology40, 745–746 (1970).
Melendez, L. V., Daniel, M. D., Hunt, R. D., Garci, F. G.: An apparently new herpesvirus from primary kidney cultures of the squirrel monkey(Saimiri sciureus). Lab. Animal Care18, 374–381 (1968).
Nazerian, K., Lindahl, T., Klein, G., Lee, L. F.: Deoxyribonucleic acid of Marek's disease virus in virus induced tumors. J. Virol.12, 841–846 (1973).
Yamamoto, K. R., Alberts, B. M.: Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology40, 734–744 (1970).
Yang, S. S., Ablashi, D., Armstrong, G., Ting, R. C.: RNA-dependent DBA polymerase associated with a simian lymphoid cell line derived from a Herpesvirus saimiri-induced lymphoma. Int. J. Can.13, 82–90 (1974).
Author information
Authors and Affiliations
Additional information
With 6 Figures
Rights and permissions
About this article
Cite this article
Simonds, J.A., Robey, W.G., Graham, B.J. et al. Purification of Herpesvirus saimiri and properties of the viral DNA. Archives of Virology 49, 249–259 (1975). https://doi.org/10.1007/BF01317543
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01317543