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. 1967 May;15(3):582–589. doi: 10.1128/am.15.3.582-589.1967

Purification of the Moloney and Rauscher Murine Leukemia Viruses by Use of Zonal Ultracentrifuge Systems

I Toplin 1
PMCID: PMC546977  PMID: 6035050

Abstract

The B-IV and B-IX zonal ultracentrifuge rotors were applied to the concentration and purification of the Moloney and Rauscher murine leukemia viruses from large volumes of infected tissue culture fluids and animal materials. Potassium tartrate, potassium citrate and sucrose gradients were used to obtain viral concentrates from the density 1.16 to 1.18 zone. Proteolytic enzyme digestion of tissue culture preparations prior to zonal ultracentrifuge processing was effective in releasing virus from cell debris and producing highly purified, though nonleukemogenic, viral concentrates. Infected Rauscher mouse plasma was processed to give highly purified infectious virus fractions. A single centrifugation of crude Rauscher mouse spleen homogenates resulted in partially purified infectious concentrates with high virus particle counts.

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Selected References

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  1. ANDERSON N. G., BARRINGER H. P., BABELAY E. F., FISHER W. D. THE B-IV ZONAL ULTRACENTRIFUGE. Life Sci. 1964 Jul;3:667–671. doi: 10.1016/0024-3205(64)90016-5. [DOI] [PubMed] [Google Scholar]
  2. ANDERSON N. G., BURGER C. L. Separation of cell components in the zonal ultracentrifuge. Science. 1962 May 18;136(3516):646–648. doi: 10.1126/science.136.3516.646. [DOI] [PubMed] [Google Scholar]
  3. ANDERSON N. G. VIRUS ISOLATION IN THE ZONAL ULTRACENTRIFUGE. Nature. 1963 Sep 21;199:1166–1168. doi: 10.1038/1991166a0. [DOI] [PubMed] [Google Scholar]
  4. CHIRIGOS M. A. STUDIES WITH THE MURINE LEUKEMOGENIC RAUSCHER VIRUS. 3. AN IN VIVO ASSAY FOR ANTI-VIRAL AGENTS. Cancer Res. 1964 Jul;24:1035–1041. [PubMed] [Google Scholar]
  5. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  6. MANAKER R. A., JENSEN E. M., KOROL W. LONG-TERM PROPAGATION OF A MURINE LEUKEMIA VIRUS IN AN ESTABLISHED CELL LINE. J Natl Cancer Inst. 1964 Aug;33:363–371. [PubMed] [Google Scholar]
  7. MCCREA J. F., EPSTEIN R. S., BARRY W. H. Use of potassium tartrate for equilibrium density-gradient centrifugation of animal viruses. Nature. 1961 Jan 21;189:220–221. doi: 10.1038/189220a0. [DOI] [PubMed] [Google Scholar]
  8. MOLONEY J. B. Biological studies on a lymphoid-leukemia virus extracted from sarcoma 37. I. Origin and introductory investigations. J Natl Cancer Inst. 1960 Apr;24:933–951. [PubMed] [Google Scholar]
  9. O'CONNOR T. E., RAUSCHER F. J., ZEIGEL R. F. DENSITY GRADIENT CENTRIFUGATION OF A MURINE LEUKEMIA VIRUS. Science. 1964 May 29;144(3622):1144–1147. doi: 10.1126/science.144.3622.1144. [DOI] [PubMed] [Google Scholar]
  10. RAUSCHER F. J. A virus-induced disease of mice characterized by erythrocytopoiesis and lymphoid leukemia. J Natl Cancer Inst. 1962 Sep;29:515–543. [PubMed] [Google Scholar]
  11. Reimer C. B., Baker R. S., Newlin T. E., Havens M. L. Influenza virus purification with the zonal ultracentrifuge. Science. 1966 Jun 3;152(3727):1379–1381. doi: 10.1126/science.152.3727.1379. [DOI] [PubMed] [Google Scholar]
  12. Sibal L. R., Fink M. A., Vice J. L., Brandt B. L., O'Connor T. E. Hemagglutination studies of the viral antigen in a murine leukemia (Rauscher). Proc Soc Exp Biol Med. 1966 Jun;122(2):591–596. doi: 10.3181/00379727-122-31199. [DOI] [PubMed] [Google Scholar]
  13. Toplin I., Riccardo D., Jensen E. M. Large-scale production of Moloney murine leukemia virus in tissue culture. Cancer. 1965 Oct;18(10):1377–1384. doi: 10.1002/1097-0142(196510)18:10<1377::aid-cncr2820181023>3.0.co;2-4. [DOI] [PubMed] [Google Scholar]
  14. Wright B. S., Lasfargues J. C. Long-term propagation of the Rauscher murine leukemia virus in tissue culture. J Natl Cancer Inst. 1965 Aug;35(2):319–327. [PubMed] [Google Scholar]

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