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. 1971 Jun;7(6):830–835. doi: 10.1128/jvi.7.6.830-835.1971

Linear, Single-Stranded Deoxyribonucleic Acid Isolated from Kilham Rat Virus

Lois Ann Salzman 1, Wesley L White 1, Tsuyoshi Kakefuda 1
PMCID: PMC356203  PMID: 4327590

Abstract

Kilham rat virus (KRV) was grown in a rat nephroma cell line and was purified by two isopycnic centrifugations in cesium chloride. The virus contains single-stranded deoxyribonucleic acid (DNA) with a molecular weight of approximately 1.6 × 106. The DNA was extracted from the virion by both phenol extraction and by 2% sodium dodecyl sulfate at 50 C. KRV DNA, extracted by both procedures, was observed in an electron microscope by using a cytochrome c or diethylaminoethyldextran monolayer. The DNA was also exposed to exonuclease I, an enzyme which hydrolyzes specifically linear, single-stranded DNA. Hydrolysis of 70 to 80% of the DNA was observed. Both the enzymatic and the electron microscope studies support the conclusion that extracted KRV DNA is a single-stranded, linear molecule. The length of the DNA was measured in the electron microscope and determined to be 1.505 ± 0.206 μm.

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Selected References

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  1. Crawford L. V., Follett E. A., Burdon M. G., McGeoch D. J. The DNA of a minute virus of mice. J Gen Virol. 1969 Jan;4(1):37–46. doi: 10.1099/0022-1317-4-1-37. [DOI] [PubMed] [Google Scholar]
  2. FIERS W., SINSHEIMER R. L. The structure of the DNA of bacteriophage phi-X174. I. The action of exopolynucleotidases. J Mol Biol. 1962 Oct;5:408–419. doi: 10.1016/s0022-2836(62)80029-1. [DOI] [PubMed] [Google Scholar]
  3. FIERS W., SINSHEIMER R. L. The structure of the DNA of bacteriophage phi-X174. III. Ultracentrifugal evidence for a ring structure. J Mol Biol. 1962 Oct;5:424–434. doi: 10.1016/s0022-2836(62)80031-x. [DOI] [PubMed] [Google Scholar]
  4. Freifelder D., Kleinschmidt A. K. Single-strand breaks in duplex DNA of coliphage T7 as demonstrated by electron microscopy. J Mol Biol. 1965 Nov;14(1):271–278. doi: 10.1016/s0022-2836(65)80246-7. [DOI] [PubMed] [Google Scholar]
  5. KILHAM L., OLIVIER L. J. A latent virus of rats isolated in tissue culture. Virology. 1959 Apr;7(4):428–437. doi: 10.1016/0042-6822(59)90071-6. [DOI] [PubMed] [Google Scholar]
  6. KLEINSCHMIDT A. K., LANG D., JACHERTS D., ZAHN R. K. [Preparation and length measurements of the total desoxyribonucleic acid content of T2 bacteriophages]. Biochim Biophys Acta. 1962 Dec 31;61:857–864. [PubMed] [Google Scholar]
  7. LEHMAN I. R., NUSSBAUM A. L. THE DEOXYRIBONUCLEASES OF ESCHERICHIA COLI. V. ON THE SPECIFICITY OF EXONUCLEASE I (PHOSPHODIESTERASE). J Biol Chem. 1964 Aug;239:2628–2636. [PubMed] [Google Scholar]
  8. LEHMAN I. R. The deoxyribonucleases of Escherichia coli. I. Purification and properties of a phosphodiesterase. J Biol Chem. 1960 May;235:1479–1487. [PubMed] [Google Scholar]
  9. PATTERSON M. S., GREENE R. C. MEASUREMENT OF LOW ENERGY BETA-EMITTERS IN AQUEOUS SOLUTION BY LIQUID SCINTILLATION COUNTING OF EMULSIONS. Anal Chem. 1965 Jun;37:854–857. doi: 10.1021/ac60226a017. [DOI] [PubMed] [Google Scholar]
  10. Robinson D. M., Hetrick F. M. Single-stranded DNA from the Kilham rat virus. J Gen Virol. 1969 Mar;4(2):269–281. doi: 10.1099/0022-1317-4-2-269. [DOI] [PubMed] [Google Scholar]
  11. Salzman L. A., Jori L. A. Characterization of the Kilham rat virus. J Virol. 1970 Feb;5(2):114–122. doi: 10.1128/jvi.5.2.114-122.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Salzman L. A., White W. L. Structural proteins of Kilham rat virus. Biochem Biophys Res Commun. 1970 Dec 24;41(6):1551–1556. doi: 10.1016/0006-291x(70)90564-4. [DOI] [PubMed] [Google Scholar]
  13. Usategui-Gomez M., Toolan H. W., Ledinko N., al-Lami F., Hopkins M. S. Single-stranded DNA from the Parvovirus, H-1. Virology. 1969 Nov;39(3):617–621. doi: 10.1016/0042-6822(69)90117-2. [DOI] [PubMed] [Google Scholar]

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