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. 1978 Sep;27(3):784–790. doi: 10.1128/jvi.27.3.784-790.1978

In vitro synthesis of double-stranded DNA from the Kilham rat virus single-stranded DNA genome.

L A Salzman, P Fabisch, R Parr, C Garon, T Wali
PMCID: PMC525866  PMID: 212593

Abstract

Double-stranded, full-length linear DNA was synthesized in vitro by using single-stranded linear DNA as a self-priming template from the parvovirus Kilham rat virus and Escherichia coli DNA polymerase "large fragment" as the polymerizing enzyme. To ascertain the order of the synthesis of the cleavage fragments and to assess the accuracy of the in vitro synthesis, restriction endonuclease cleavage sites with known recognition sequences were mapped on the DNA. Comparing the cleavage pattern of the synthesized DNA with that of double-stranded viral DNA isolated from infected cells confirms that the in vitro synthesis produces a faithful copy of the viral single-stranded genome. Electron micrographs of the in vitro product reveal it to be a double-stranded linear molecule.

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Selected References

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  1. Allet B., Jeppesen P. G., Katagiri K. J., Delius H. Mapping the DNA fragments produced by cleavage by lambda DNA with endonuclease RI. Nature. 1973 Jan 12;241(5385):120–123. doi: 10.1038/241120a0. [DOI] [PubMed] [Google Scholar]
  2. Bourguignon G. J., Tattersall P. J., Ward D. C. DNA of minute virus of mice: self-priming, nonpermuted, single-stranded genome with a 5'-terminal hairpin duplex. J Virol. 1976 Oct;20(1):290–306. doi: 10.1128/jvi.20.1.290-306.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cavalier-Smith T. Palindromic base sequences and replication of eukaryote chromosome ends. Nature. 1974 Aug 9;250(5466):467–470. doi: 10.1038/250467a0. [DOI] [PubMed] [Google Scholar]
  4. Danna K. J., Sack G. H., 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. doi: 10.1016/0022-2836(73)90122-8. [DOI] [PubMed] [Google Scholar]
  5. Gunther M., May P. Isolation and structural characterization of monomeric and dimeric forms of replicative intermediates of Kilham rat virus DNA. J Virol. 1976 Oct;20(1):86–95. doi: 10.1128/jvi.20.1.86-95.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Iyer V. N., Rupp W. D. Usefulness of benzoylated naphthoylated DEAE-cellulose to distinguish and fractionate double-stranded DNA bearing different extents of single-stranded regions. Biochim Biophys Acta. 1971 Jan 1;228(1):117–126. doi: 10.1016/0005-2787(71)90551-x. [DOI] [PubMed] [Google Scholar]
  8. Lavelle G., Li A. T. Isolation of intracellular replicative forms and progeny single strands of DNA from parvovirus KRV in sucrose gradients containing guanidine hydrochloride. Virology. 1977 Jan;76(1):464–467. doi: 10.1016/0042-6822(77)90324-5. [DOI] [PubMed] [Google Scholar]
  9. Modrich P., Zabel D. EcoRI endonuclease. Physical and catalytic properties of the homogenous enzyme. J Biol Chem. 1976 Oct 10;251(19):5866–5874. [PubMed] [Google Scholar]
  10. Nathans D., Smith H. O. Restriction endonucleases in the analysis and restructuring of dna molecules. Annu Rev Biochem. 1975;44:273–293. doi: 10.1146/annurev.bi.44.070175.001421. [DOI] [PubMed] [Google Scholar]
  11. Rhode S. L., 3rd Replication process of the parvovirus H-1. VI. Characterization of a replication terminus of H-1 replicative-form DNA. J Virol. 1977 Feb;21(2):694–712. doi: 10.1128/jvi.21.2.694-712.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Salzman L. A., Jori L. A. Characterization of the Kilham rat virus. J Virol. 1970 Feb;5(2):114–122. doi: 10.1128/jvi.5.2.114-122.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Salzman L. A., White W. L., Kakefuda T. Linear, single-stranded deoxyribonucleic acid isolated from Kilham rat virus. J Virol. 1971 Jun;7(6):830–835. doi: 10.1128/jvi.7.6.830-835.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Salzman L. A., White W. L. Structural proteins of Kilham rat virus. Biochem Biophys Res Commun. 1970 Dec 24;41(6):1551–1556. doi: 10.1016/0006-291x(70)90564-4. [DOI] [PubMed] [Google Scholar]
  15. Salzman L. A., White W. In vivo conversion of the single-stranded DNA of the kilham rat virus to a double-stranded form. J Virol. 1973 Feb;11(2):299–305. doi: 10.1128/jvi.11.2.299-305.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sedat J. W., Kelly R. B., Sinsheimer R. L. Fractionation of nucleic acid on benzoylated-naphthoylated DEAE cellulose. J Mol Biol. 1967 Jun 28;26(3):537–540. doi: 10.1016/0022-2836(67)90321-x. [DOI] [PubMed] [Google Scholar]
  17. Setlow P., Brutlag D., Kornberg A. Deoxyribonucleic acid polymerase: two distinct enzymes in one polypeptide. I. A proteolytic fragment containing the polymerase and 3' leads to 5' exonuclease functions. J Biol Chem. 1972 Jan 10;247(1):224–231. [PubMed] [Google Scholar]
  18. Straus S. E., Sebring E. D., Rose J. A. Concatemers of alternating plus and minus strands are intermediates in adenovirus-associated virus DNA synthesis. Proc Natl Acad Sci U S A. 1976 Mar;73(3):742–746. doi: 10.1073/pnas.73.3.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tattersall P., Ward D. C. Rolling hairpin model for replication of parvovirus and linear chromosomal DNA. Nature. 1976 Sep 9;263(5573):106–109. doi: 10.1038/263106a0. [DOI] [PubMed] [Google Scholar]

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