Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1967 Apr;1(2):302–307. doi: 10.1128/jvi.1.2.302-307.1967

Purification and Properties of Reovirus Ribonucleic Acid

W J Iglewski 1, R M Franklin 1
PMCID: PMC375228  PMID: 4318949

Abstract

NaClO4 was employed in a technique for the rapid extraction of reovirus ribonucleic acid (RNA). The extracted RNA, which was purified in a Cs2SO4 equilibrium density gradient, had a buoyant density of 1.61 g/cm3 and a sedimentation coefficient of 15S in a 7 to 20% sucrose gradient. It was 90% resistant to ribonuclease in a solution of high ionic strength (0.1 m NaCl). The sensitivity of reovirus RNA to ribonuclease increased with decreasing ionic strength. The thermal denaturation transition of the RNA began at 78 C and was complete at 85 C. The Tm of the transition was 81 C in 0.01 m tris(hydroxymethyl)aminomethane buffer (pH 7.2) containing 0.001 m ethylenediaminetetraacetate. Thermal denaturation of reovirus RNA resulted in the formation of three ribonuclease-sensitive fractions. Denaturation at 25 C in the presence of dimethyl sulfoxide resulted in the formation of two ribonuclease-sensitive fractions.

Full text

PDF
307

Selected References

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

  1. BURGI E., HERSHEY A. D. Sedimentation rate as a measure of molecular weight of DNA. Biophys J. 1963 Jul;3:309–321. doi: 10.1016/s0006-3495(63)86823-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Franklin R. M. Replication of bacteriophage ribonucleic acid: some physical properties of single-stranded, double-stranded, and branched viral ribonucleic acid. J Virol. 1967 Feb;1(1):64–75. doi: 10.1128/jvi.1.1.64-75.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Freifelder D. A rapid technique for the preparation of purified bacteriophage DNA or RNA from crude lysates. Biochim Biophys Acta. 1965 Oct 11;108(2):318–319. doi: 10.1016/0005-2787(65)90020-1. [DOI] [PubMed] [Google Scholar]
  4. Freifelder D. Effect of NaCIO-4 on bacteriophage: release of DNA and evidence for population heterogeneity. Virology. 1966 Apr;28(4):742–750. doi: 10.1016/0042-6822(66)90258-3. [DOI] [PubMed] [Google Scholar]
  5. GOMATOS P. J., STOECKENIUS W. ELECTRON MICROSCOPE STUDIES ON REOVIRUS RNA. Proc Natl Acad Sci U S A. 1964 Dec;52:1449–1455. doi: 10.1073/pnas.52.6.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gomatos P. J., Tamm I. THE SECONDARY STRUCTURE OF REOVIRUS RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):707–714. doi: 10.1073/pnas.49.5.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Katz L., Penman S. The solvent denaturation of double-stranded RNA from poliovirus infected HeLa cells. Biochem Biophys Res Commun. 1966 May 25;23(4):557–560. doi: 10.1016/0006-291x(66)90765-0. [DOI] [PubMed] [Google Scholar]
  8. Kudo H., Graham A. F. Synthesis of reovirus ribonucleic acid in L cells. J Bacteriol. 1965 Oct;90(4):936–945. doi: 10.1128/jb.90.4.936-945.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LERNER A. M. CONCENTRATION AND PURIFICATION OF VIRUSES WITH SPECIAL REFERENCE TO REOVIRUSES. Bacteriol Rev. 1964 Dec;28:391–396. doi: 10.1128/br.28.4.391-396.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Loh P. C., Soergel M. Growth characteristics of reovirus type 2: actinomycin D and the synthesis of viral RNA. Proc Natl Acad Sci U S A. 1965 Sep;54(3):857–863. doi: 10.1073/pnas.54.3.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Shatkin A. J. Inactivity of purified reovirus RNA as a template for E. coli polymerases in vitro. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1721–1728. doi: 10.1073/pnas.54.6.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES