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. 1979 Dec;38(6):1132–1139. doi: 10.1128/aem.38.6.1132-1139.1979

In vitro production of Rauscher murine leukemia virus: influence of culture age on biological properties.

O R Robinson Jr, A Perry, G P Shibley, M de Maio
PMCID: PMC291257  PMID: 93428

Abstract

Large-scale production and concentration procedures have been standardized to study the biological properties of Rauscher leukemia virus produced from the high-passaged JLS-V9-H mouse bone marrow cell line. Virus produced early (days 4 to 6) in the harvest and refeed cycle contained higher levels of ribonucleic acid-directed deoxyribonucleic acid polymerase activity and was more infectious than Rauscher leukemia virus produced later (days 7 to 10) in the growth period. The peak of virus production as detected by physical assays (virus particle count, protein, and p30 antigen) was highest at day 6, whereas the optimum biological and ribonucleic acid-directed deoxyribonucleic acid polymerase activity occurred 24 h earlier. When product characterization values of each concentrate were adjusted to a specific activity (i.e., per milligram of protein) basis, virus particle counts averaged 4 x 10(11) through days 5 to 9, and the peak infectivity occurred at day 4, whereas ribonucleic acid-directed deoxyribonucleic acid polymerase activity was highest at day 4 (endogenous) and 5 (exogenous). Sodium dodecyl sulfate-polyacrylamide gel analysis revealed only slight differences in the polypeptide pattern of Rauscher leukemia virus harvested from cultures of varying age, although Rauscher leukemia virus produced between days 3 and 5 contained more glycoprotein than either earlier or later harvests.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AXELRAD A. A., STEEVES R. A. ASSAY FOR FRIEND LEUKEMIA VIRUS: RAPID QUANTITATIVE METHOD BASED ON ENUMERATION OF MACROSCOPIC SPLEEN FOCI IN MICE. Virology. 1964 Nov;24:513–518. doi: 10.1016/0042-6822(64)90199-0. [DOI] [PubMed] [Google Scholar]
  2. Aaronson S. A., Stephenson J. R., Greenberger J. S. Cellular replication and the persistence of inducible RNA type C viruses. J Virol. 1974 Jun;13(6):1404–1407. doi: 10.1128/jvi.13.6.1404-1407.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aaronson S. A., Stephenson J. R. Independent segregation of loci for activation of biologically distinguishable RNA C-type viruses in mouse cells. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2055–2058. doi: 10.1073/pnas.70.7.2055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Aaronson S. A., Todaro G. J., Scolnick E. M. Induction of murine C-type viruses from clonal lines of virus-free BALB-3T3 cells. Science. 1971 Oct 8;174(4005):157–159. doi: 10.1126/science.174.4005.157. [DOI] [PubMed] [Google Scholar]
  5. August J. T., Bolognesi D. P., Fleissner E., Gilden R. V., Nowinski R. C. A proposed nomenclature for the virion proteins of oncogenic RNA viruses. Virology. 1974 Aug;60(2):595–600. doi: 10.1016/0042-6822(74)90356-0. [DOI] [PubMed] [Google Scholar]
  6. Bader J. P., Steck T. L. Analysis of the ribonucleic acid of murine leukemia virus. J Virol. 1969 Oct;4(4):454–459. doi: 10.1128/jvi.4.4.454-459.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Canaani E., Duesberg P. Role of subunits of 60 to 70S avian tumor virus ribonucleic acid in its template activity for the viral deoxyribonucleic acid polymerase. J Virol. 1972 Jul;10(1):23–31. doi: 10.1128/jvi.10.1.23-31.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chattopadhyay S. K., Lowy D. R., Teich N. M., Levine A. S., Rowe W. P. Evidence that the AKR murine-leukemia-virus genome is complete in DNA of the high-virus AKR mouse and incomplete in the DNA of the "virus-negative" NIH mouse. Proc Natl Acad Sci U S A. 1974 Jan;71(1):167–171. doi: 10.1073/pnas.71.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Collins M. J., Jr, Parker J. C. Murine virus contaminants of leukemia viruses and transplantable tumors. J Natl Cancer Inst. 1972 Oct;49(4):1139–1143. [PubMed] [Google Scholar]
  10. Eckner R. J., Steeves R. A. A classification of the murine leukemia viruses. Neutralization of pseudotypes of Friend spleen focus-forming virus by type-specific murine antisera. J Exp Med. 1972 Oct 1;136(4):832–850. doi: 10.1084/jem.136.4.832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gazdar A. F., Russell E. K., Minna J. D. Replication of mouse-tropic and xenotropic strains of murine leukemia virus in human x mouse hybrid cells. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2642–2645. doi: 10.1073/pnas.71.7.2642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gelb L. D., Milstien J. B., Martin M. A., Aaronson S. A. Characterization of murine leukaemia virus-specific DNA present in normal mouse cells. Nat New Biol. 1973 Jul 18;244(133):76–79. doi: 10.1038/newbio244076a0. [DOI] [PubMed] [Google Scholar]
  13. Hodge H. M., Klein F., Bandyopadhyay A. K., Robinson O. R., Jr, Shibley G. P. Adaptation and infection of mouse bone marrow (JLS-V9) cells in suspension culture for production of Rauscher leukemia virus. Appl Microbiol. 1974 Jan;27(1):224–231. doi: 10.1128/am.27.1.224-231.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnson R. W., Perry A., Robinson O. R., Jr, Shibley G. P. Method for reproducible large-volume production and purification of Rauscher murine leukemia virus. Appl Environ Microbiol. 1976 Feb;31(2):182–188. doi: 10.1128/aem.31.2.182-188.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Lowy D. R., Rowe W. P., Teich N., Hartley J. W. Murine leukemia virus: high-frequency activation in vitro by 5-iododeoxyuridine and 5-bromodeoxyuridine. Science. 1971 Oct 8;174(4005):155–156. doi: 10.1126/science.174.4005.155. [DOI] [PubMed] [Google Scholar]
  17. Monroe J. H., Brandt P. M. Rapid semiquantitative method for screening large numbers of virus samples by negative staining electron microscopy. Appl Microbiol. 1970 Aug;20(2):259–262. doi: 10.1128/am.20.2.259-262.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. PLUZNIK D. H., SACHS L. QUANTITATION OF A MURINE LEUKEMIA VIRUS WITH A SPLEEN COLONY ASSAY. J Natl Cancer Inst. 1964 Sep;33:535–546. [PubMed] [Google Scholar]
  19. Perry A., Robinson O. R., Jr, Shibley G. P. Enhanced production of Rauscher leukemia virus after infection of high-passaged JLS-V9 cells. Appl Environ Microbiol. 1978 Sep;36(3):526–529. doi: 10.1128/aem.36.3.526-529.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scolnick E., Rands E., Aaronson S. A., Todaro G. J. RNA-dependent DNA polymerase activity in five RNA viruses: divalent cation requirements. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1789–1796. doi: 10.1073/pnas.67.4.1789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smith R. E. High specific infectivity avian RNA tumor viruses. Virology. 1974 Aug;60(2):543–547. doi: 10.1016/0042-6822(74)90348-1. [DOI] [PubMed] [Google Scholar]
  22. Steeves R. A., Eckner R. J., Bennett M., Mirand E. A., Trudel P. J. Isolation and characterization of a lymphatic leukemia virus in the Friend virus complex. J Natl Cancer Inst. 1971 Jun;46(6):1209–1217. [PubMed] [Google Scholar]
  23. Stephension J. R., Reynolds R. K., Tronick S. R., Aaronson S. A. Distribution of three classes of endogenous type-C RNA viruses among inbred strains of mice. Virology. 1975 Oct;67(2):404–414. doi: 10.1016/0042-6822(75)90442-0. [DOI] [PubMed] [Google Scholar]
  24. Stephenson J. R., Aaronson S. A. Segregation of loci for C-type virus induction in strains of mice with high and low incidence of leukemia. Science. 1973 May 25;180(4088):865–866. doi: 10.1126/science.180.4088.865. [DOI] [PubMed] [Google Scholar]
  25. Syrewicz J. J., Naso R. B., Wang C. S., Arlinghaus R. B. Purification of large amounts of murine ribonucleic acid tumor viruses produced in roller bottle cultures. Appl Microbiol. 1972 Sep;24(3):488–494. doi: 10.1128/am.24.3.488-494.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Toplin I., Sottong P. Large-volume purification of tumor viruses by use of zonal centrifuges. Appl Microbiol. 1972 May;23(5):1010–1014. doi: 10.1128/am.23.5.1010-1014.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Watson J. D. The structure and assembly of murine leukemia virus: intracellular viral RNA. Virology. 1971 Sep;45(3):586–597. doi: 10.1016/0042-6822(71)90174-7. [DOI] [PubMed] [Google Scholar]
  29. Wright B. S., Lasfargues J. C. Attenuation of the Rauscher murine leukemia virus through serial passages in tissue culture. Natl Cancer Inst Monogr. 1966 Sep;22:685–700. [PubMed] [Google Scholar]
  30. Wright B. S., O'Brien P. A., Shibley G. P., Mayyasi S. A., Lasfargues J. C. Infection of an established mouse bone marrow cell line (JLS-V9) with Rauscher and Moloney murine leukemia viruses. Cancer Res. 1967 Sep;27(9):1672–1677. [PubMed] [Google Scholar]

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