Linear, Single-Stranded Deoxyribonucleic Acid Isolated from Kilham Rat Virus |
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Authors: | Lois Ann Salzman Wesley L. White Tsuyoshi Kakefuda |
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Affiliation: | 1Laboratory of Biology of Viruses, National Institute of Allergy and Infectious Diseases, and Chemistry Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014 |
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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 x 10(6). 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 mum. |
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