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Specific Origin in SV40 DNA Replication

Nature New Biology volume 236, pages 200–202 (1972)Cite this article

Abstract

THE oncogenic virus SV40 contains a covalently closed circular DNA molecule of 3 × 106 molecular weight1. In infected permissive cells, SV40 DNA replicates through a Cairns type intermediate2 with the parental strands forming a partially twisted, covalently closed molecule3. We have used a specific bacterial restriction endonuclease4 to analyse SV40 DNA replication. The restriction endonuclease of H. influenzae makes double-strand breaks in DNA at specific hexanucleotide sequences5, 6 and splits SV40 DNA into eleven fragments, separable by Polyacrylamide gel electrophoresis, ranging in molecular weight from 6.5 × 105 (about 20% of the molecule) to 7.4 × 104 (about 2.5% of the molecule). The largest eight fragments are present in the digest in amounts equimolar with the starting DNA4. Therefore, by digesting labelled replicating SV40 DNA and newly completed DNA, and measuring the relative yield of each fragment, we could determine whether a particular region of the DNA is synthesized first or last and also estimate the time needed to replicate one molecule completely.

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Authors and Affiliations

  1. Department of Microbiology, Johns Hopkins University School of Medicine, Baltimore, Maryland, 21205

    DANIEL NATHANS & KATHLEEN J. DANNA

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  1. DANIEL NATHANS
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  2. KATHLEEN J. DANNA
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NATHANS, D., DANNA, K. Specific Origin in SV40 DNA Replication. Nature New Biology 236, 200–202 (1972). https://doi.org/10.1038/newbio236200a0

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