Abstract
Procedures are described for production and purification of large amounts of Rous sarcoma virus. The virus was produced by Rous sarcoma virus-transformed chicken embryo fibroblasts in roller culture which produced up to 6 mg of virus per day per liter of supernatant fluid. Various methods of concentrating virus were evaluated; pelleting yielded the best results in terms of recovery of infectious virus. Purification was achieved by means of successive velocity and equilibrium density centrifugation by using sucrose solutions made in low-salt buffer. A rapid method for the optical density measurement of virus concentration was also developed.
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Selected References
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- BALUDA M. A., GOETZ I. E. Morphological conversion of cell cultures by avian myeloblastosis virus. Virology. 1961 Oct;15:185–199. doi: 10.1016/0042-6822(61)90234-3. [DOI] [PubMed] [Google Scholar]
- BONAR R. A., BEARD J. W. Virus of avian myeloblastosis. XII. Chemical constitution. J Natl Cancer Inst. 1959 Jul;23(1):183–197. [PubMed] [Google Scholar]
- Bader J. P. A method for the propagation of large amounts of Rous sarcoma virus. Virology. 1968 Sep;36(1):140–142. doi: 10.1016/0042-6822(68)90124-4. [DOI] [PubMed] [Google Scholar]
- Bolognesi D. P., Bauer H. Polypeptides of avian RNA tumor viruses. 1. Isolation and physical and chemical analysis. Virology. 1970 Dec;42(4):1097–1112. doi: 10.1016/0042-6822(70)90357-0. [DOI] [PubMed] [Google Scholar]
- Cheung K. S., Smith R. E., Stone M. P., Joklik W. K. Comparison of immature (rapid harvest) and mature Rous sarcoma virus particles. Virology. 1972 Dec;50(3):851–864. doi: 10.1016/0042-6822(72)90439-4. [DOI] [PubMed] [Google Scholar]
- Day E. D., McMillan P. N., Mickey D. D., Appel S. H. Zonal centrifuge profiles of rat brain homogenates: instability in sucrose, stability in iso-osmotic Ficoll-sucrose. Anal Biochem. 1971 Jan;39(1):29–45. doi: 10.1016/0003-2697(71)90458-1. [DOI] [PubMed] [Google Scholar]
- Duesberg P. H., Robinson H. L., Robinson W. S., Huebner R. J., Turner H. C. Proteins of Rous sarcoma virus. Virology. 1968 Sep;36(1):73–86. doi: 10.1016/0042-6822(68)90118-9. [DOI] [PubMed] [Google Scholar]
- Duff R. G., Vogt P. K. Characteristics of two new avian tumor virus subgroups. Virology. 1969 Sep;39(1):18–30. doi: 10.1016/0042-6822(69)90344-4. [DOI] [PubMed] [Google Scholar]
- Graf T. A plaque assay for avian RNA tumor viruses. Virology. 1972 Nov;50(2):567–578. doi: 10.1016/0042-6822(72)90408-4. [DOI] [PubMed] [Google Scholar]
- Graf T., Bauer H., Gelderblom H., Bolognesi D. P. Studies on the reproductive and cell-converting abilities of avian sarcoma viruses. Virology. 1971 Feb;43(2):427–441. doi: 10.1016/0042-6822(71)90315-1. [DOI] [PubMed] [Google Scholar]
- HAM R. G. An improved nutrient solution for diploid Chinese hamster and human cell lines. Exp Cell Res. 1963 Feb;29:515–526. doi: 10.1016/s0014-4827(63)80014-2. [DOI] [PubMed] [Google Scholar]
- JOKLIK W. K. The purification fo four strains of poxvirus. Virology. 1962 Sep;18:9–18. doi: 10.1016/0042-6822(62)90172-1. [DOI] [PubMed] [Google Scholar]
- Kawai S., Hanafusa H. Plaque assay for some strains of avian leukosis virus. Virology. 1972 Apr;48(1):126–135. doi: 10.1016/0042-6822(72)90120-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Luftig R. B., Kilham S. S. An electron microscope study of Rauscher leukemia virus. Virology. 1971 Nov;46(2):277–297. doi: 10.1016/0042-6822(71)90030-4. [DOI] [PubMed] [Google Scholar]
- Macieira-Coelho A. Relationship between DNA synthesis and cell density in normal and virus-transformed cells. Int J Cancer. 1967 Jul 15;2(4):297–303. doi: 10.1002/ijc.2910020404. [DOI] [PubMed] [Google Scholar]
- McSharry J., Benzinger R. Concentration and purification of vesicular stomatitis virus by polyethylene glycol "precipitation". Virology. 1970 Mar;40(3):745–746. doi: 10.1016/0042-6822(70)90219-9. [DOI] [PubMed] [Google Scholar]
- Moscovici C. A quantitative assay for avian myeloblastosis virus. Proc Soc Exp Biol Med. 1967 Aug-Sep;125(4):1213–1215. doi: 10.3181/00379727-125-32316. [DOI] [PubMed] [Google Scholar]
- Moscovici C., Vogt P. K. Effects of genetic cellular resistance on cell transformation and virus replication in chicken hematopoietic cell cultures infected with avian myeloblastosis virus (BAI-A). Virology. 1968 Aug;35(4):487–497. doi: 10.1016/0042-6822(68)90278-x. [DOI] [PubMed] [Google Scholar]
- RUBIN H., CORNELIUS A., FANSHIER L. The pattern of congenital transmission of an avian lekosis virus. Proc Natl Acad Sci U S A. 1961 Jul 15;47:1058–1069. doi: 10.1073/pnas.47.7.1058. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RUBIN H., VOGT P. K. An avian leukosis virus associated with stocks of Rous sarcoma virus. Virology. 1962 May;17:184–194. doi: 10.1016/0042-6822(62)90096-x. [DOI] [PubMed] [Google Scholar]
- Smith R. E., Moscovici C. The oncogenic effects of nontransforming viruses from avian myeloblastosis virus. Cancer Res. 1969 Jul;29(7):1356–1366. [PubMed] [Google Scholar]
- Smith R. E., Zweerink H. J., Joklik W. K. Polypeptide components of virions, top component and cores of reovirus type 3. Virology. 1969 Dec;39(4):791–810. doi: 10.1016/0042-6822(69)90017-8. [DOI] [PubMed] [Google Scholar]
- Spear P. G., Roizman B. Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpesvirion. J Virol. 1972 Jan;9(1):143–159. doi: 10.1128/jvi.9.1.143-159.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stromberg K., Litwack M. D., Desmukes B. Structural studies on avian myeloblastosis virus: rapid purification and quantitation. Proc Soc Exp Biol Med. 1972 Oct;141(1):215–221. doi: 10.3181/00379727-141-36745. [DOI] [PubMed] [Google Scholar]
- Temin H. M. Studies on carcinogenesis by avian sarcoma viruses. 3. The differential effect of serum and polyanions on multiplication of uninfected and converted cells. J Natl Cancer Inst. 1966 Aug;37(2):167–175. [PubMed] [Google Scholar]
- Toyoshima K., Vogt P. K. Enhancement and inhibition of avian sarcoma viruses by polycations and polyanions. Virology. 1969 Jul;38(3):414–426. doi: 10.1016/0042-6822(69)90154-8. [DOI] [PubMed] [Google Scholar]
- VOGT P. K., RUBIN H. Studies on the assay and multiplication of avian myeloblastosis virus. Virology. 1963 Jan;19:92–104. doi: 10.1016/0042-6822(63)90029-1. [DOI] [PubMed] [Google Scholar]