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. 1977 Sep;23(3):827–832. doi: 10.1128/jvi.23.3.827-832.1977

Analysis of polyoma virus DNA replicative intermediates by agarose gel electrophoresis.

R F Martin
PMCID: PMC515896  PMID: 197274

Abstract

Agarose gel electrophoresis has been used to fractionate polyoma virus DNA replicative intermediates (RI) according to maturity. Approximate electrophoretic mobility versus maturity relationships were obtained for both intact (supercoiled) and nicked (relaxed) RI. There was considerable overlap between the supercoiled and relaxed RI populations after electrophoretic fractionation. Intact RI could be recovered from preparative agarose gels for further analysis by centrifugation, electron microscopy, re-electrophoresis, or nuclease digestion.

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Selected References

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  1. Bourgaux P., Bourgaux-Ramoisy D. Unwinding of replicating polyoma virus DNA. J Mol Biol. 1972 Oct 14;70(3):399–413. doi: 10.1016/0022-2836(72)90548-7. [DOI] [PubMed] [Google Scholar]
  2. Dingman C. W., Fisher M. P., Kakefuda T. Role of molecular conformation in determining the electrophoretic properties of polynucleotides in agarose-acrylamide gels. II. Biochemistry. 1972 Mar 28;11(7):1242–1250. doi: 10.1021/bi00757a020. [DOI] [PubMed] [Google Scholar]
  3. Flint D. H., Harrington R. E. Gel electrophoresis of deoxyribonucleic acid. Biochemistry. 1972 Dec 5;11(25):4858–4864. doi: 10.1021/bi00775a034. [DOI] [PubMed] [Google Scholar]
  4. Griffin B. E., Fried M., Cowie A. Polyoma DNA: a physical map. Proc Natl Acad Sci U S A. 1974 May;71(5):2077–2081. doi: 10.1073/pnas.71.5.2077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  6. Keller W., Wendel I. Stepwise relaxation of supercoiled SV40 DNA. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):199–208. doi: 10.1101/sqb.1974.039.01.026. [DOI] [PubMed] [Google Scholar]
  7. Magnusson G., Craig R., Närkhammar M., Reichard P., Staub M., Warner H. Replication of polyoma DNA: effects of hydroxyurea and arabinosyl cytosine. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):227–233. doi: 10.1101/sqb.1974.039.01.029. [DOI] [PubMed] [Google Scholar]
  8. Magnusson G. Hydroxyurea-induced accumulation of short fragments during polyoma DNA replication. I. Characterization of fragments. J Virol. 1973 Sep;12(3):600–608. doi: 10.1128/jvi.12.3.600-608.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Magnusson G., Pigiet V., Winnacker E. L., Abrams R., Reichard P. RNA-linked short DNA fragments during polyoma replication. Proc Natl Acad Sci U S A. 1973 Feb;70(2):412–415. doi: 10.1073/pnas.70.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mayer A., Levine A. J. DNA replication in SV40-infected cells. 8. The distribution of replicating molecules at different stages of replication in SV40-infected cells. Virology. 1972 Nov;50(2):328–338. doi: 10.1016/0042-6822(72)90384-4. [DOI] [PubMed] [Google Scholar]
  11. Otto B., Reichard P. Replication of polyoma DNA in isolated nuclei. V. Complementation of in vitro DNA replication. J Virol. 1975 Feb;15(2):259–267. doi: 10.1128/jvi.15.2.259-267.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pettersson U., Mulder C., Deluis H., Sharp P. A. Cleavage of adenovirus type 2 DNA into six unique fragments by endonuclease R-RI. Proc Natl Acad Sci U S A. 1973 Jan;70(1):200–204. doi: 10.1073/pnas.70.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pigiet V., Eliasson R., Reichard P. Replication of polyoma DNA in isolated nuclei. 3. The nucleotide sequence at the RNA-DNA junction of nascent strands. J Mol Biol. 1974 Mar 25;84(1):197–216. doi: 10.1016/0022-2836(74)90222-8. [DOI] [PubMed] [Google Scholar]
  14. Pigiet V., Winnacker E. L., Eliasson R., Reichard P. Discontinuous elongation of both strands at the replication forks in polyoma DNA replication. Nat New Biol. 1973 Oct 17;245(146):203–205. doi: 10.1038/newbio245203a0. [DOI] [PubMed] [Google Scholar]
  15. STUDIER F. W. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA. J Mol Biol. 1965 Feb;11:373–390. doi: 10.1016/s0022-2836(65)80064-x. [DOI] [PubMed] [Google Scholar]
  16. Sharp P. A., 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. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  17. Tegtmeyer P., Macasaet F. Simian virus 40 deoxyribonucleic acid synthesis: analysis by gel electrophoresis. J Virol. 1972 Oct;10(4):599–604. doi: 10.1128/jvi.10.4.599-604.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Weinberg R. A., Loening U., Willems M., Penman S. Acrylamide gel electrophoresis of HeLa cell nucleolar RNA. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1088–1095. doi: 10.1073/pnas.58.3.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Winnacker E. L., Magnusson G., Reichard P. Replication of polyoma DNA in isolated nuclei. I. Characterization of the system from mouse fibroblast 3T6 cells. J Mol Biol. 1972 Dec 30;72(3):523–537. doi: 10.1016/0022-2836(72)90172-6. [DOI] [PubMed] [Google Scholar]

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