Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1982 Aug;43(2):566–573. doi: 10.1128/jvi.43.2.566-573.1982

New physical map of bacteriophage T5 DNA.

M Rhoades
PMCID: PMC256160  PMID: 6287031

Abstract

The locations of 103 cleavage sites, produced by 13 restriction endonucleases, were mapped on the DNA of bacteriophage T5. Single- and double-digest fragment sizes were determined by agarose gel electrophoresis, using restriction fragments of phi X174 DNA and lambda DNA as molecular weight standards. Map coordinates were determined by a computer-based least-squares procedures (J. Schroeder and F. Blattner, Gene [Amst] 4:167-174, 1978). The fragment sizes predicted by the final map are all within 2% of the measured values. Based on this analysis, T5st(+) DNA contains 121,300 base pairs (Mr, 80.3 X 10(6) and has a terminal repetition of 10,160 base pairs (Mr, 6.7 X 10(6)). Restriction endonuclease analysis after treatment with exonuclease III and a single-strand-specific endonuclease allowed precise localization of five of the natural single-chain interruptions in T5 DNA. Revised locations for several T5 deletions were also determined.

Full text

PDF
573

Images in this article

Selected References

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

  1. Brunel F., Davison J., Merchez M. Cloning of bacteriophage T5 DNA fragments in plasmid pBR322 and bacteriophage lambda gtWES. Gene. 1979 Dec;8(1):53–68. doi: 10.1016/0378-1119(79)90007-6. [DOI] [PubMed] [Google Scholar]
  2. Brunel F., Davison J. Restriction insensitivity in bacteriophage T5. III. Characterization of EcoRI-sensitive mutants by restriction analysis. J Mol Biol. 1979 Mar 15;128(4):527–543. doi: 10.1016/0022-2836(79)90291-2. [DOI] [PubMed] [Google Scholar]
  3. Chen M. J., Locker J., Weiss S. B. The physical mapping of bacteriophage T5 transfer tRNAs. J Biol Chem. 1976 Jan 25;251(2):536–547. [PubMed] [Google Scholar]
  4. Daniels D. L., de Wet J. R., Blattner F. R. New map of bacteriophage lambda DNA. J Virol. 1980 Jan;33(1):390–400. doi: 10.1128/jvi.33.1.390-400.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. 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]
  8. Hunt C., Desai S. M., Vaughan J., Weiss S. B. Bacteriophage T5 transfer RNA. Isolation and characterization of tRNA species and refinement of the tRNA gene map. J Biol Chem. 1980 Apr 10;255(7):3164–3173. [PubMed] [Google Scholar]
  9. Lanni Y. T. First-step-transfer deoxyribonucleic acid of bacteriophage T5. Bacteriol Rev. 1968 Sep;32(3):227–242. doi: 10.1128/br.32.3.227-242.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nichols B. P., Donelson J. E. Mapping and cloning of Eco RI-fragments of bacteriophage T5+ DNA. Nucleic Acids Res. 1977 Nov;4(11):3715–3726. doi: 10.1093/nar/4.11.3715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. 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]
  14. Rhoades M., Schwartz J., Wahl J. M. New deletion mutant of bacteriophage T5. J Virol. 1980 Nov;36(2):622–626. doi: 10.1128/jvi.36.2.622-626.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rogers S. G., Godwin E. A., Shinosky E. S., Rhoades M. Interruption-deficient mutants of bacteriophage T5 I. Isolation and general properties. J Virol. 1979 Feb;29(2):716–725. doi: 10.1128/jvi.29.2.716-725.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rogers S. G., Hamlett N. V., Rhoades M. Interruption-deficient mutants of bacteriophage T5. II. Properties of a mutant lacking a specific interruption. J Virol. 1979 Feb;29(2):726–734. doi: 10.1128/jvi.29.2.726-734.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rubenstein I. Heat-stable mutants of T5 phage. I. The physical properties of the phage and their DNA molecules. Virology. 1968 Nov;36(3):356–376. doi: 10.1016/0042-6822(68)90161-x. [DOI] [PubMed] [Google Scholar]
  18. Sanger F., Coulson A. R., Friedmann T., Air G. M., Barrell B. G., Brown N. L., Fiddes J. C., Hutchison C. A., 3rd, Slocombe P. M., Smith M. The nucleotide sequence of bacteriophage phiX174. J Mol Biol. 1978 Oct 25;125(2):225–246. doi: 10.1016/0022-2836(78)90346-7. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Scheible P. P., Rhoades M. Heteroduplex mapping of heat-resistant deletion mutants of bacteriophage t5. J Virol. 1975 May;15(5):1276–1280. doi: 10.1128/jvi.15.5.1276-1280.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schroeder J. L., Blattner F. R. Least-squares method for restriction mapping. Gene. 1978 Oct;4(2):167–174. doi: 10.1016/0378-1119(78)90028-8. [DOI] [PubMed] [Google Scholar]
  22. Stüber D., Bujard H. Electron microscopy of DNA: determination of absolute molecular weights and linear density. Mol Gen Genet. 1977 Sep 9;154(3):299–303. doi: 10.1007/BF00571286. [DOI] [PubMed] [Google Scholar]
  23. Stüber D., Delius H., Bujard H. Electron microscopic analysis of in vitro transcriptional complexes: mapping of promoters of the coliphage T5 genome. Mol Gen Genet. 1978 Oct 30;166(2):141–149. doi: 10.1007/BF00285916. [DOI] [PubMed] [Google Scholar]
  24. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  25. Tchernov A. P., Kouzmin N. P., Fodor I. Physical mapping of cleavage sites recognized by restriction endonucleases on the genome of bacteriophage T5. Gene. 1978 Jul;3(4):293–302. doi: 10.1016/0378-1119(78)90039-2. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. von Gabain A., Bujard H. Interaction of Escherichia coli RNA polymerase with promoters of several coliphage and plasmid DNAs. Proc Natl Acad Sci U S A. 1979 Jan;76(1):189–193. doi: 10.1073/pnas.76.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. von Gabain A., Hayward G. S., Bujard H. Physical mapping of the HindIII, EcoRI, Sal and Sma restriction endonuclease cleavage fragments from bacteriophage T5 DNA. Mol Gen Genet. 1976 Feb 2;143(3):279–290. doi: 10.1007/BF00269404. [DOI] [PubMed] [Google Scholar]

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

RESOURCES