Physical and Biological Properties of Phage φ29 Deoxyribonucleic Acid (original) (raw)

Abstract

Deoxyribonucleic acid (DNA) molecules having a mean length of 5.8 μm were released from purified Bacillus subtilis bacteriophage φ29 with 2 m sodium perchlorate. Small 0.1 to 0.2-μm molecules were also detected in these DNA preparations. Since intact single chains annealed to form linear duplex molecules, phage φ29 DNA was found to be nonpermuted. The molecular weights of single chains of φ29 DNA were approximately half that of native DNA, as determined by analytical band sedimentation in CsCl, indicating that φ29 DNA is composed of two continuous polynucleotide chains. The molecular weight values of native and annealed φ29 DNA from sedimentation agreed with the molecular weight values obtained from electron microscopy. The infectivity of φ29 DNA was reduced to a low level by alkaline denaturation and was partially restored by annealing.

1185

Images in this article

Selected References

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

  1. Anderson D. L., Hickman D. D., Reilly B. E. Structure of Bacillus subtilis bacteriophage phi 29 and the length of phi 29 deoxyribonucleic acid. J Bacteriol. 1966 May;91(5):2081–2089. doi: 10.1128/jb.91.5.2081-2089.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bott K. F., Wilson G. A. Development of competence in the Bacillus subtilis transformation system. J Bacteriol. 1967 Sep;94(3):562–570. doi: 10.1128/jb.94.3.562-570.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bruner R., Vinograd J. The evaluation of standard sedimentation coefficients of sodium RNA and sodium DNA from sedimentation velocity data in concentrated NaCl and CsCl solutions. Biochim Biophys Acta. 1965 Sep 6;108(1):18–29. doi: 10.1016/0005-2787(65)90104-8. [DOI] [PubMed] [Google Scholar]
  4. Freifelder D. Effect of NaCIO-4 on bacteriophage: release of DNA and evidence for population heterogeneity. Virology. 1966 Apr;28(4):742–750. doi: 10.1016/0042-6822(66)90258-3. [DOI] [PubMed] [Google Scholar]
  5. KLEINSCHMIDT A. K., LANG D., JACHERTS D., ZAHN R. K. [Preparation and length measurements of the total desoxyribonucleic acid content of T2 bacteriophages]. Biochim Biophys Acta. 1962 Dec 31;61:857–864. [PubMed] [Google Scholar]
  6. Lang D., Bujard H., Wolff B., Russell D. Electron microscopy of size and shape of viral DNA in solutions of different ionic strengths. J Mol Biol. 1967 Jan 28;23(2):163–181. doi: 10.1016/s0022-2836(67)80024-x. [DOI] [PubMed] [Google Scholar]
  7. MacHattie L. A., Ritchie D. A., Thomas C. A., Jr, Richardson C. C. Terminal repetition in permuted T2 bacteriophage DNA molecules. J Mol Biol. 1967 Feb 14;23(3):355–363. doi: 10.1016/s0022-2836(67)80110-4. [DOI] [PubMed] [Google Scholar]
  8. REILLY B. E., SPIZIZEN J. BACTERIOPHAGE DEOXYRIBONUCLEATE INFECTION OF COMPETENT BACILLUS SUBTILIS. J Bacteriol. 1965 Mar;89:782–790. doi: 10.1128/jb.89.3.782-790.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ritchie D. A., Thomas C. A., Jr, MacHattie L. A., Wensink P. C. Terminal repetition in non-permuted T3 and T7 bacteriophage DNA molecules. J Mol Biol. 1967 Feb 14;23(3):365–376. doi: 10.1016/s0022-2836(67)80111-6. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. THOMAS C. A., Jr, MACHATTIE L. A. CIRCULAR T2 DNA MOLECULES. Proc Natl Acad Sci U S A. 1964 Nov;52:1297–1301. doi: 10.1073/pnas.52.5.1297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. VINOGRAD J., BRUNER R., KENT R., WEIGLE J. Band-centrifugation of macromolecules and viruses in self-generating density gradients. Proc Natl Acad Sci U S A. 1963 Jun;49:902–910. doi: 10.1073/pnas.49.6.902. [DOI] [PMC free article] [PubMed] [Google Scholar]