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J Virol. 1971 June; 7(6): 830-835
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

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

Lois Ann Salzman, Wesley L. White and Tsuyoshi Kakefuda

1 Laboratory of Biology of Viruses, National Institute of Allergy and Infectious Diseases, and Chemistry Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014

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 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.


J Virol. 1971 June; 7(6): 830-835
Copyright © 1971 American Society for Microbiology. All Rights Reserved.







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