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. 1979 Feb;29(2):517–522. doi: 10.1128/jvi.29.2.517-522.1979

Reverse transcriptase from avian myeloblastosis virus.

G E Houts, M Miyagi, C Ellis, D Beard, J W Beard
PMCID: PMC353184  PMID: 85721

Abstract

From lots of 20 to 30 g of avian myeloblastosis virus RNA-dependent DNA polymerase was obtained in preparations of purity greater than 95% by using a two-step column chromatographic procedure employing DEAE (DE 52) and carboxymethylcellulose (CM 52.). Yields of RNA-dependent DNA polymerase varied from approximately 20,000 to 35,000 U/g of virus. Specific activity of the enzyme was about 35,000 to 60,000 U/mg of protein. Free of detectable RNase activity, the product exhibited a molecular weight of about 160,000, an isoelectric point of 6.5, and approximately 2 mol of fatty acid per mol of enzyme.

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Selected References

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  1. BEAUDREAU G. S., BECKER C. Virus of avian myeloblastosis. X. Photometric microdetermination of adenosinetriphosphatase activity. J Natl Cancer Inst. 1958 Feb;20(2):339–349. [PubMed] [Google Scholar]
  2. Bonar R. A., Sverak L., Bolognesi D. P., Langlois A. J., Beard D., Beard J. W. Ribonucleic acid components of BAI strain A (myeloblastosis) avian tumor virus. Cancer Res. 1967 Jun;27(6):1138–1157. [PubMed] [Google Scholar]
  3. Devos R., Van Emmelo J., Celen P., Gillis E., Fiers W. Synthesis by avian-myeloblastosis-virus RNA-dependent DNA polymerase of discrete reverse transcripts of bacteriophage RNA polyadenylated in vitro. Eur J Biochem. 1977 Oct 3;79(2):419–432. doi: 10.1111/j.1432-1033.1977.tb11824.x. [DOI] [PubMed] [Google Scholar]
  4. ECKERT E. A., BEARD D., BEARD J. W. Dose-response relations in experimental transmission of avian erythromyeloblastic leukosis. I. Host-response to the virus. J Natl Cancer Inst. 1951 Oct;12(2):447–463. [PubMed] [Google Scholar]
  5. ECKERT E. A., BEARD D., BEARD J. W. Dose-response relations in experimental transmission of avian erythromyeoloblastic leukosis. V. Influence of host age and route of virus inoculation. J Natl Cancer Inst. 1955 Apr;15(5):1195–1207. [PubMed] [Google Scholar]
  6. ECKERT E. A., WATERS N. F., BURMESTER B. R., BEARD D., BEARD J. W. Dose-response relations in experimental transmission of avian erythromyeloblastic leukosis. IV. Strain differences in host-response to the virus. J Natl Cancer Inst. 1954 Apr;14(5):1067–1080. [PubMed] [Google Scholar]
  7. Efstratiadis A., Maniatis T., Kafatos F. C., Jeffrey A., Vournakis J. N. Full length and discrete partial reverse transcripts of globin and chorion mRNAs. Cell. 1975 Apr;4(4):367–378. doi: 10.1016/0092-8674(75)90157-9. [DOI] [PubMed] [Google Scholar]
  8. Faras A. J., Taylor J. M., McDonnell J. P., Levinson W. E., Bishop J. M. Purification and characterization of the deoxyribonucleic acid polymerase associated with Rous sarcoma virus. Biochemistry. 1972 Jun 6;11(12):2334–2342. doi: 10.1021/bi00762a020. [DOI] [PubMed] [Google Scholar]
  9. Friedman E. Y., Rosbash M. The syntheiss of high yields of full-length reverse transcripts of globin mRNA. Nucleic Acids Res. 1977 Oct;4(10):3455–3471. doi: 10.1093/nar/4.10.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grandgenett D. P. Purification of the alpha subunit of avian myeloblastosis virus DNA polymerase by polyuridylic acid-sepharose. J Virol. 1976 Oct;20(1):348–350. doi: 10.1128/jvi.20.1.348-350.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Higuchi R., Paddock G. V., Wall R., Salser W. A general method for cloning eukaryotic structural gene sequences. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3146–3150. doi: 10.1073/pnas.73.9.3146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Houts G. E., Miyagi M., Ellis C., Beard D., Watson K. F., Beard J. W. Protein kinase from avian myeloblastosis virus. J Virol. 1978 Feb;25(2):546–552. doi: 10.1128/jvi.25.2.546-552.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hurwitz J., Leis J. P. RNA-dependent DNA polymerase activity of RNA tumor viruses. I. Directing influence of DNA in the reaction. J Virol. 1972 Jan;9(1):116–129. doi: 10.1128/jvi.9.1.116-129.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kedes L. H., Chang A. C., Houseman D., Cohen S. N. Isolation of histone genes from unfractionated sea urchin DNA by subculture cloning in E. coli. Nature. 1975 Jun 12;255(5509):533–538. doi: 10.1038/255533a0. [DOI] [PubMed] [Google Scholar]
  15. Modak M. J., Marcus S. L. Purification and properties of Rauscher leukemia virus DNA polymerase and selective inhibition of mammalian viral reverse transcriptase by inorganic phosphate. J Biol Chem. 1977 Jan 10;252(1):11–19. [PubMed] [Google Scholar]
  16. Monahan J. J., Harris S. E., Woo S. L., Robberson D. L., O'Malley B. W. The synthesis and properties of the complete complementary DNA transcript of ovalbumin mRNA. Biochemistry. 1976 Jan 13;15(1):223–233. doi: 10.1021/bi00646a034. [DOI] [PubMed] [Google Scholar]
  17. Myers J. C., Spiegelman S., Kacian D. L. Synthesis of full-length DNA copies of avian myeloblastosis virus RNA in high yields. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2840–2843. doi: 10.1073/pnas.74.7.2840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rabbitts T. H. Bacterial cloning of plasmids carrying copies of rabbit globin messenger RNA. Nature. 1976 Mar 18;260(5548):221–225. doi: 10.1038/260221a0. [DOI] [PubMed] [Google Scholar]
  19. SHARP D. G., BEARD J. W. Virus of avian erythromyeloblastic leukosis. IV. Sedimentation, density and hydration. Biochim Biophys Acta. 1954 May;14(1):12–17. doi: 10.1016/0006-3002(54)90124-9. [DOI] [PubMed] [Google Scholar]

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