1887

Abstract

Summary

By combining the techniques of acid precipitation, organic solvent extraction, enzyme treatment and differential centrifugation, concentration of encephalomyocarditis virus more than 1000-fold with 30% to 50% yield and a degree of purification of more than 1 × 10 was achieved. Yield was estimated by adding virus labelled with P to crude virus and measuring the percentage recovery both of the radioactive label and of the total number of plaque forming units in the final product. The degree of purification was determined by adding both P-and H-labelled, uninfected host-cell material to crude virus and measuring the extent to which the final product was contaminated with radioactive label. Pure virus comprised a homogeneous preparation of polyhedral particles with a particle to plaque forming units ratio of 250 ± 40 and gave a single, symmetrical peak in the analytical ultracentrifuge.

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/content/journal/jgv/10.1099/0022-1317-5-2-291
1969-09-01
2024-04-25
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References

  1. Bellett A. J. D., Burness A. T. H. (1963); Intracellular sites of synthesis of encephalomyocarditis virus components in Krebs-2 ascites tumour cells. J. gen. Microbiol 30:131
    [Google Scholar]
  2. Bruno G. A., Christian J. E. (1961); Determination of carbon-14 in aqueous bicarbonate solutions by liquid scintillation counting techniques. Anal. Chem 33:1216
    [Google Scholar]
  3. Burness A. T. H. (1967); Separation of plaque-type variants of encephalomyocarditis virus by chromatography on calcium phosphate. J. Virol 1:308
    [Google Scholar]
  4. Burness A. T. H., Moss J. (1967); Use of kieselguhr to increase cell production for animal virus investigations. Nature. Lond. 213:833
    [Google Scholar]
  5. Burness A. T. H., Vizoso A. D. (1961); The relationship between molecular size of ribonucleic acid and its chromatographic profile on calcium phosphate. Biochim. biophys. Acta 49:225
    [Google Scholar]
  6. Burness A. T. H., Walter D. S. (1967); Protein components of encephalomyocarditis virus. Nature, Lond 215:1350
    [Google Scholar]
  7. Burness A. T. H., Vizoso A. D., Clothier F. W. (1963); Sedimentation coefficients of encephalo- myocarditis virus and its ribonucleic acid. Nature. Lond 197:1177
    [Google Scholar]
  8. Dische Z. (1955); Color reactions of nucleic acid components. In The Nucleic Acids 1285 Chargaff E., Davidson J. N. New York: Academic Press;
    [Google Scholar]
  9. Faulkner P., Martin E. M., Sved S., Valentine R. C., Work T. S. (1961); Studies on protein and nucleic acid metabolism in virus-infected mammalian cells. 2. The isolation, crystallization and chemical characterization of mouse encephalomyocarditis virus. Biochem. J. 80:597
    [Google Scholar]
  10. Gessler A. E., Bender C. E., Parkinson M. C. (1956); A new and rapid method for isolating viruses by selective fluorocarbon deproteinization. Trans. N.Y. Acad. Sci. 18:701
    [Google Scholar]
  11. Hoskins J. M., Sanders F. K. (1957); Propagation of mouse encephalomyocarditis virus in ascites tumour cells maintained in vitro. Br. J. exp. Path. 38:268
    [Google Scholar]
  12. Kaighn M. E., Moscarello M. A., Fuerst C. R. (1964); Purification of murine encephalomyocarditis virus. Virology 23:183
    [Google Scholar]
  13. Kitano T., Haruna I., Watanabe I. (1961); Purification and concentration of virus by an organic solvent system. Virology 15:503
    [Google Scholar]
  14. Krug R. M., Franklin R. M. (1964); Studies on the time of synthesis of Mengovirus ribonucleic acid and protein. Virology 22:48
    [Google Scholar]
  15. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. (1951); Protein measurement with the folin phenol reagent. J. biol. Chem 193:265
    [Google Scholar]
  16. Markham R. (1967); The ultracentrifuge. In Methods in Virology II1 Maramorosch K., Koprowski H. New York and London: Academic Press;
    [Google Scholar]
  17. Martin E. M., Malec J., Sved S., Work T. S. (1961); Studies on protein and nucleic acid metabolism in virus-infected mammalian cells. I. Encephalomyocarditis virus in Krebs II mouse ascites tumour cells. Biochem. J. 80:585
    [Google Scholar]
  18. Mellema J. E., Van Bruggen E. F. J., Gruber M. (1968); An assessment of negative staining in the electron microscopy of low molecular weignt proteins. J. molec. Biol. 31:75
    [Google Scholar]
  19. Montagnier L., Sanders F. K. (1963); Sedimentation properties of infective ribonucleic acid extracted from encephalomyocarditis virus. Nature, Lond. 197:1178
    [Google Scholar]
  20. Moscarello M. A., Kaighn M. E. (1964); The amino acid composition of three plaque-type mutants of encephalomyocarditis virus. Biochim. biophys. Acta 90:161
    [Google Scholar]
  21. Philipson L., Albertsson P. A., Frick G. (1960); The purification and concentration of viruses by aqueous polymer phase systems. Virology 11:553
    [Google Scholar]
  22. Roizman B., Mayer M. M., Rapp H. J. (1958); Immunochemical studies of poliovirus. III. Further studies on the immunologic and physical properties of poliovirus particles produced in tissue culture. J. Immunol. 81:419
    [Google Scholar]
  23. Rueckert R. R., Schäfer W. (1965); Studies on the structure of viruses of the Columbia SK Group. I. Purification and properties of ME virus grown in Ehrlich ascites cell suspensions. Virology 26:333
    [Google Scholar]
  24. Sanders F. K., Huppert J., Hoskins J. M. (1958); Replication of an animal virus. Symp. Soc. exp. Biol. 12:123
    [Google Scholar]
  25. Scharff M. D., Levintow L. (1963); Quantitative study of the formation of poliovirus antigens in infected HeLa cells. Virology 19:491
    [Google Scholar]
  26. Taverne J., Marshall J. H., Fulton F. (1958); The purification and concentration of viruses and virus soluble antigens on calcium phosphate. Nature, Lond 184:1655
    [Google Scholar]
  27. Volkin E., Cohn W. E. (1954); Estimation of nucleic acids. In Methods of Biochemical Analysis I287 Glick D. New York: Interscience Publishers;
    [Google Scholar]
  28. Wheatley D. N. (1964); Influence of various substances on production of ascites tumour. Nature, Lond 202:1348
    [Google Scholar]
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