Polyoma DNA: a physical map.
PDF (1.1M)
Journal: 1974/August - Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
PUBMED: 4365585
Abstract:
The action of restriction enzymes on polyoma DNA was studied with uniformly (32)P-labeled viral DNA obtained from either infected 3T6 or secondary mouse-embryo cells. Three restriction enzymes were used to construct a physical map of the polyoma genome. An enzyme from Hemophilus parainfluenzae, Hpa(II), cleaved polyma DNA into eight unique fragments (Hpa(II)-1 to Hpa(II)-8), ranging in size from 27.3 to 1.8% of the genome. An enzyme from Hemophilus influenzae, Hin(III), gave two fragments (56 and 44%); and a third enzyme from Escherichia coli, EcoR(I), cut at a single unique site. The physical map of the polyma genome was constructed from methods involving: (1) further digestion of the fragments produced by enzymes EcoR(I) and Hin(III) with Hpa(II), and (2) analysis of the products of partial digestion with Hpa(II). Analysis by electron microscopy of replicating DNA molecules (less than 50% replicated) cut with the Hin(III) enzyme, in combination with other studies, has indicated that the origin of DNA replication is located at 71 +/- 3 map units from the EcoR(I) cleavage site, probably in Hpa(II)-5.
Open in
PUBMED | PMC | Google Scholar | Wikipedia
Relations:
Content
Citations
(104)
References
(12)
Drugs
(1)
Chemicals
(6)
Organisms
(4)
Processes
(4)
Similar articles
Articles by the same authors
Discussion board
Proc Natl Acad Sci U S A 71(5): 2077-2081
PMID: 4365585

Polyma DNA: A Physical Map

Abstract

The action of restriction enzymes on polyoma DNA was studied with uniformly P-labeled viral DNA obtained from either infected 3T6 or secondary mouse-embryo cells. Three restriction enzymes were used to construct a physical map of the polyoma genome. An enzyme from Hemophilus parainfluenzae, HpaII, cleaved polyma DNA into eight unique fragments (HpaII-1 to HpaII-8), ranging in size from 27.3 to 1.8% of the genome. An enzyme from Hemophilus influenzae, HinIII, gave two fragments (56 and 44%); and a third enzyme from Escherichia coli, EcoRI, cut at a single unique site. The physical map of the polyma genome was constructed from methods involving: (1) further digestion of the fragments produced by enzymes EcoRI and HinIII with HpaII, and (2) analysis of the products of partial digestion with HpaII.

Analysis by electron microscopy of replicating DNA molecules (less than 50% replicated) cut with the HinIII enzyme, in combination with other studies, has indicated that the origin of DNA replication is located at 71 ± 3 map units from the EcoRI cleavage site, probably in HpaII-5.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
icon of scanned page 2077
2077
icon of scanned page 2078
2078
icon of scanned page 2079
2079
icon of scanned page 2080
2080
icon of scanned page 2081
2081

Images in this article

Image
Image
on p.2078
Image
Image
on p.2079
Image
Image
on p.2080
Image
Image
on p.2080
Image
Image
on p.2080
Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. [PubMed] [Google Scholar]
  • Radloff R, Bauer W, Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521.[PMC free article] [PubMed] [Google Scholar]
  • Sharp PA, Sugden B, Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. [PubMed] [Google Scholar]
  • Danna KJ, Sack GH, Jr, Nathans D. Studies of simian virus 40 DNA. VII. A cleavage map of the SV40 genome. J Mol Biol. 1973 Aug 5;78(2):363–376. [PubMed] [Google Scholar]
  • Huang ES, Newbold JE, Pagano JS. Analysis of simian virus 40 DNA with the restriction enzyme of Haemophilus aegyptius, endonuclease Z. J Virol. 1973 Apr;11(4):508–514.[PMC free article] [PubMed] [Google Scholar]
  • Salzman NP, Sebring ED, Radonovich M. Unwinding of parental strands during simian virus 40 DNA replication. J Virol. 1973 Oct;12(4):669–676.[PMC free article] [PubMed] [Google Scholar]
  • Crawford LV, Syrett C, Wilde A. The replication of polyoma DNA. J Gen Virol. 1973 Dec;21(3):515–521. [PubMed] [Google Scholar]
  • Brockman WW, Lee TN, Nathans D. The evolution of new species of viral DNA during serial passage of simian virus 40 at high multiplicity. Virology. 1973 Aug;54(2):384–397. [PubMed] [Google Scholar]
  • Danna KJ, Nathans D. Bidirectional replication of Simian Virus 40 DNA. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3097–3100.[PMC free article] [PubMed] [Google Scholar]
  • Nathans D, Danna KJ. Studies of SV40 DNA. 3. Differences in DNA from various strains of SV40. J Mol Biol. 1972 Mar 14;64(2):515–518. [PubMed] [Google Scholar]
  • Khoury G, Martin MA, Lee TN, Danna KJ, Nathans D. A map of simian virus 40 transcription sites expressed in productively infected cells. J Mol Biol. 1973 Aug 5;78(2):377–389. [PubMed] [Google Scholar]
  • Chen CY, Hutchison CA, 3rd, Edgell MH. Isolation and genetic localization of three phi-X174 promoter regions. Nat New Biol. 1973 Jun 20;243(129):233–236. [PubMed] [Google Scholar]
Department of Molecular Virology, Imperial Cancer Research Fund, P.O. Box 123, Lincoln's Inn Fields, London WC2A 3PX, England
Department of Tumor Virus Genetics, Imperial Cancer Research Fund, P.O. Box 123, Lincoln's Inn Fields, London WC2A 3PX, England
Copyright notice
Abstract
The action of restriction enzymes on polyoma DNA was studied with uniformly P-labeled viral DNA obtained from either infected 3T6 or secondary mouse-embryo cells. Three restriction enzymes were used to construct a physical map of the polyoma genome. An enzyme from Hemophilus parainfluenzae, HpaII, cleaved polyma DNA into eight unique fragments (HpaII-1 to HpaII-8), ranging in size from 27.3 to 1.8% of the genome. An enzyme from Hemophilus influenzae, HinIII, gave two fragments (56 and 44%); and a third enzyme from Escherichia coli, EcoRI, cut at a single unique site. The physical map of the polyma genome was constructed from methods involving: (1) further digestion of the fragments produced by enzymes EcoRI and HinIII with HpaII, and (2) analysis of the products of partial digestion with HpaII.Analysis by electron microscopy of replicating DNA molecules (less than 50% replicated) cut with the HinIII enzyme, in combination with other studies, has indicated that the origin of DNA replication is located at 71 ± 3 map units from the EcoRI cleavage site, probably in HpaII-5.
Collaboration tool especially designed for Life Science professionals.Drag-and-drop any entity to your messages.