Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1979 Jun;30(3):777–786. doi: 10.1128/jvi.30.3.777-786.1979

Physical map of bacteriophage BF23 DNA: terminal redundancy and localization of single-chain interruptions.

M Rhoades, B Lange-Gustafson
PMCID: PMC353388  PMID: 158099

Abstract

The DNA of bacteriophage BF23 possesses two structural features, localized single-chain interruptions and a large terminal repetition, previously described for T5, a closely related virus. As is the case for T5, single-chain interruptions occur with variable frequencies at a small number of fixed sites within one strand of the double-stranded BF23 genome. The sites where interruptions occur with the highest frequencies were napped by an electrophoretic analysis of the single-stranded fragments produced by denaturation of BF23 DNA. The positions of these fragments were determined by degrading BF23 DNA to various extents with lambda exonuclease and observing the relative order with which they were (i) degraded or (ii) released intact from the undenatured duplex. The exact locations of the interruptions were determined from analysis of analogous duplex fragments produced by degrading exonuclease III-treated BF23 DNA with a single-strand-specific endonuclease. BF23 has five principal sites (located at 7.9, 18.7, 32.4, 65.8, and 99.6% from the left end of the DNA) where interruptions occur in most molecules. The principal interruptions in T5 DNA occur at similar positions. The locations of eight secondary interruptions in BF23 DNA were also determined. In general, BF23 DNA has fewer secondary interruptions than t5 dna, although there is at least one location where an interruption occurs with a greater frequency in BF23. The presence of a terminal repetition in BF23 DNA was demonstrated by annealing ligase-repaired molecules that had been partially digested with lambda exonuclease. If the complementary sequences at both ends of the DNA were exposed by exonuclease treatment, the duplex segment that resulted from annealing could be released by digestion with a single-strand-specific endonuclease. This segment was analyzed by agarose gel electrophoresis and found to represent 8.4% of BF23 DNA.

Full text

PDF
777

Images in this article

Selected References

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

  1. Carter D. M., Radding C. M. The role of exonuclease and beta protein of phage lambda in genetic recombination. II. Substrate specificity and the mode of action of lambda exonuclease. J Biol Chem. 1971 Apr 25;246(8):2502–2512. [PubMed] [Google Scholar]
  2. Dürwald H., Hoffmann-Berling H. Endonuclease-I-deficient and ribonuclease I-deficient Escherichia coli mutants. J Mol Biol. 1968 Jul 14;34(2):331–346. doi: 10.1016/0022-2836(68)90257-x. [DOI] [PubMed] [Google Scholar]
  3. HERTEL R., MARCHI L., MULLER K. Density mutants of phage T5. Virology. 1962 Dec;18:576–581. doi: 10.1016/0042-6822(62)90060-0. [DOI] [PubMed] [Google Scholar]
  4. Hamlett N. V., Lange-Gufstafson B., Rhoades M. Physical map of the bacteriophage T5 genome based on the cleavage products of the restriction endonucleases SalI, SmaI, BamI, and HpaI. J Virol. 1977 Oct;24(1):249–260. doi: 10.1128/jvi.24.1.249-260.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hayward G. S., Smith M. G. The chromosome of bacteriophage T5. I. Analysis of the single-stranded DNA fragments by agarose gel electrophoresis. J Mol Biol. 1972 Feb 14;63(3):383–395. doi: 10.1016/0022-2836(72)90435-4. [DOI] [PubMed] [Google Scholar]
  6. Hayward G. S. Unique double-stranded fragments of bacteriophage T5 DNA resulting from preferential shear-induced breakage at nicks. Proc Natl Acad Sci U S A. 1974 May;71(5):2108–2112. doi: 10.1073/pnas.71.5.2108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jacquemin-Sablon A., Richardson C. C. Analysis of the interruptions in bacteriophage T5 DNA. J Mol Biol. 1970 Feb 14;47(3):477–493. doi: 10.1016/0022-2836(70)90316-5. [DOI] [PubMed] [Google Scholar]
  8. LANG D., Shaw A. R., McCorquodale D. J. Molecular weights of DNA from bacteriophages T5, T5st(O), BF23, and BF23st(4). J Virol. 1975 Jan;17(1):296–297. doi: 10.1128/jvi.17.1.296-297.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lange-Gustafson B., Rhoades M. Physical map of bacteriophage BF23 DNA: restriction enzyme analysis. J Virol. 1979 Jun;30(3):923–928. doi: 10.1128/jvi.30.3.923-928.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Masamune Y., Fleischman R. A., Richardson C. C. Enzymatic removal and replacement of nucleotides at single strand breaks in deoxyribonucleic acid. J Biol Chem. 1971 Apr 25;246(8):2680–2691. [PubMed] [Google Scholar]
  11. McCorquodale D. J. The T-odd bacteriophages. CRC Crit Rev Microbiol. 1975 Dec;4(2):101–159. doi: 10.3109/10408417509111574. [DOI] [PubMed] [Google Scholar]
  12. Mizobuchi K., Anderson G. C., McCorquodale D. J. Abortive infection by bacteriophage BF23 due to the colicin Ib factor. I. Genetic studies of nonrestricted and amber mutants of bacteriophage BF23. Genetics. 1971 Jul;68(3):323–340. doi: 10.1093/genetics/68.3.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nisioka T., Ozeki H. Early abortive lysis by phage BF23 in Escherichia coli K-12 carrying the colicin Ib factor. J Virol. 1968 Nov;2(11):1249–1254. doi: 10.1128/jvi.2.11.1249-1254.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rhoades M. Cleavage of T5 DNA by the Escherichia coli R-I restriction endonuclease. Virology. 1975 Mar;64(1):170–179. doi: 10.1016/0042-6822(75)90089-6. [DOI] [PubMed] [Google Scholar]
  15. Rhoades M. Localization of single-chain interruptions in bacteriophage T5 DNA. II. Electrophoretic studies. J Virol. 1977 Sep;23(3):737–750. doi: 10.1128/jvi.23.3.737-750.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rhoades M., Rhoades E. A. Terminal repetition in the DNA of bacteriophage T5. J Mol Biol. 1972 Aug 21;69(2):187–200. doi: 10.1016/0022-2836(72)90224-0. [DOI] [PubMed] [Google Scholar]
  17. Rogers S. G., Rhoades M. Bacteriophage T5-induced endonucleases that introduce site-specific single-chain interruptions in duplex DNA. Proc Natl Acad Sci U S A. 1976 May;73(5):1576–1580. doi: 10.1073/pnas.73.5.1576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Scheible P. P., Rhoades E. A., Rhoades M. Localization of single-chain interruptions in bacteriophage T5 DNA I. Electron microscopic studies. J Virol. 1977 Sep;23(3):725–736. doi: 10.1128/jvi.23.3.725-736.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sriprakash K. S., Lundh N., Huh MM-O, Radding C. M. The specificity of lambda exonuclease. Interactions with single-stranded DNA. J Biol Chem. 1975 Jul 25;250(14):5438–5445. [PubMed] [Google Scholar]
  20. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES