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. 1973 Jan;70(1):200–204. doi: 10.1073/pnas.70.1.200

Cleavage of Adenovirus Type 2 DNA into Six Unique Fragments by Endonuclease R·RI

Ulf Pettersson 1, Carel Mulder 1, Hajo Delius 1, Phillip A Sharp 1
PMCID: PMC433215  PMID: 4567334

Abstract

The DNA of adenovirus type 2 was cleaved by restriction endonuclease R·RI into six fragments. These fragments were separated by electrophoresis on composite agarose-polyacrylamide gels. Their molecular weights ranged from 1.1 × 106 to 13.6 × 106, as measured by electron microscopy. Each fragment represented a unique segment of adenovirus type 2 DNA since: (i) the fragments were obtained in equimolar amounts; (ii) the sum of their molecular weights was equal to the molecular weight of complete adenovirus DNA; and (iii) each fragment exhibited a rate of renaturation that was inversely proportional to its size.

Keywords: restriction enzyme, electron microscopy, DNA gel electrophoresis, DNA sequence complexity

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Danna K., Nathans D. Specific cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus influenzae. Proc Natl Acad Sci U S A. 1971 Dec;68(12):2913–2917. doi: 10.1073/pnas.68.12.2913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Doerfler W., Kleinschmidt A. K. Denaturation pattern of the DNA of adenovirus type 2 as determined by electron microscopy. J Mol Biol. 1970 Jun 28;50(3):579–593. doi: 10.1016/0022-2836(70)90086-0. [DOI] [PubMed] [Google Scholar]
  3. Espejo R. T., Canelo E. S., Sinsheimer R. L. DNA of bacteriophage PM2: a closed circular double-stranded molecule. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1164–1168. doi: 10.1073/pnas.63.4.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Freeman A. E., Black P. H., Vanderpool E. A., Henry P. H., Austin J. B., Huebner R. J. Transformation of primary rat embryo cells by adenovirus type 2. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1205–1212. doi: 10.1073/pnas.58.3.1205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Freifelder D. Molecular weights of coliphages and coliphage DNA. IV. Molecular weights of DNA from bacteriophages T4, T5 and T7 and the general problem of determination of M. J Mol Biol. 1970 Dec 28;54(3):567–577. doi: 10.1016/0022-2836(70)90127-0. [DOI] [PubMed] [Google Scholar]
  6. GREEN M., PINA M. Biochemical studies on adenovirus multiplication. IV. Isolation, purification, and chemical analysis of adenovirus. Virology. 1963 May;20:199–207. doi: 10.1016/0042-6822(63)90157-0. [DOI] [PubMed] [Google Scholar]
  7. Garon C. F., Berry K. W., Rose J. A. A unique form of terminal redundancy in adenovirus DNA molecules. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2391–2395. doi: 10.1073/pnas.69.9.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Green M., Piña M., Kimes R., Wensink P. C., MacHattie L. A., Thomas C. A., Jr Adenovirus DNA. I. Molecular weight and conformation. Proc Natl Acad Sci U S A. 1967 May;57(5):1302–1309. doi: 10.1073/pnas.57.5.1302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hedgpeth J., Goodman H. M., Boyer H. W. DNA nucleotide sequence restricted by the RI endonuclease. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3448–3452. doi: 10.1073/pnas.69.11.3448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lonberg-Holm K., Philipson L. Early events of virus-cell interaction in an adenovirus system. J Virol. 1969 Oct;4(4):323–338. doi: 10.1128/jvi.4.4.323-338.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mertz J. E., Davis R. W. Cleavage of DNA by R 1 restriction endonuclease generates cohesive ends. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3370–3374. doi: 10.1073/pnas.69.11.3370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Morrow J. F., Berg P. Cleavage of Simian virus 40 DNA at a unique site by a bacterial restriction enzyme. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3365–3369. doi: 10.1073/pnas.69.11.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mulder C., Delius H. Specificity of the break produced by restricting endonuclease R 1 in Simian virus 40 DNA, as revealed by partial denaturation mapping. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3215–3219. doi: 10.1073/pnas.69.11.3215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Peacock A. C., Dingman C. W. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry. 1968 Feb;7(2):668–674. doi: 10.1021/bi00842a023. [DOI] [PubMed] [Google Scholar]
  15. Pettersson U., Philipson L., Höglund S. Structural proteins of adenoviruses. I. Purification and characterization of the adenovirus type 2 hexon antigen. Virology. 1967 Dec;33(4):575–590. doi: 10.1016/0042-6822(67)90057-8. [DOI] [PubMed] [Google Scholar]
  16. Prage L., Pettersson U., Höglund S., Lonberg-Holm K., Philipson L. Structural proteins of adenoviruses. IV. Sequential degradation of the adenovirus type 2 virion. Virology. 1970 Oct;42(2):341–358. doi: 10.1016/0042-6822(70)90278-3. [DOI] [PubMed] [Google Scholar]
  17. Wetmur J. G., Davidson N. Kinetics of renaturation of DNA. J Mol Biol. 1968 Feb 14;31(3):349–370. doi: 10.1016/0022-2836(68)90414-2. [DOI] [PubMed] [Google Scholar]
  18. Wolfson J., Dressler D. Adenovirus-2 DNA contains an inverted terminal repetition. Proc Natl Acad Sci U S A. 1972 Oct;69(10):3054–3057. doi: 10.1073/pnas.69.10.3054. [DOI] [PMC free article] [PubMed] [Google Scholar]

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