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. 1977 Oct;24(1):249–260. doi: 10.1128/jvi.24.1.249-260.1977

Physical map of the bacteriophage T5 genome based on the cleavage products of the restriction endonucleases SalI, SmaI, BamI, and HpaI.

N V Hamlett, B Lange-Gufstafson, M Rhoades
PMCID: PMC515927  PMID: 904024

Abstract

A physical map of the bacteriophage T5 genome was constructed by ordering the fragments produced by cleavage of T5 DNA with the restriction endonucleases SalI (4 fragments), SmaI (4 fragments), BamI (5 fragments), and HpaI (28 fragments). The following techniques were used to order the fragments. (i) Digestion of DNA from T5 heat-stable deletion mutants was used to identify fragments located in the deletable region. (ii) Fragments near the ends of the T5 DNA molecule were located by treating T5 DNA with lambda exonuclease before restriction endonuclease cleavage. (iii) Fragments spanning other restriction endonuclease cleavage sites were identified by combined digestion of T5 DNA with two restriction endonucleases. (iv) The general location of some fragments was determined by isolating individual restriction fragments from agarose gels and redigesting the isolated fragments with a second restriction enzyme. (v) Treatment of restriction digests with lambda exonuclease before digestion with a second restriction enzyme was used to identify fragments near, but not spanning, restriction cleavage sites. (vi) Exonucleases III treatment of T5 DNA before restriction endonuclease cleavage was used to locate fragments spanning or near the natural T5 single-chain interruptions. (vii) Analysis of the products of incomplete restriction endonuclease cleavage was used to identify adjacent fragments.

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Selected References

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

  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. Blin N., von Gabain A., Bujard H. Isolation of large molecular weight DNA from agarose gels for further digestion by restriction enzymes. FEBS Lett. 1975 Apr 15;53(1):84–86. doi: 10.1016/0014-5793(75)80688-0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Hayward G. S., Smith M. G. The chromosome of bacteriophage T5. II. Arrangement of the single-stranded DNA fragments in the T5 + and T5st(O) chromosomes. J Mol Biol. 1972 Feb 14;63(3):397–407. doi: 10.1016/0022-2836(72)90436-6. [DOI] [PubMed] [Google Scholar]
  5. Kelly T. J., Jr, Smith H. O. A restriction enzyme from Hemophilus influenzae. II. J Mol Biol. 1970 Jul 28;51(2):393–409. doi: 10.1016/0022-2836(70)90150-6. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Lanni Y. Functions of two genes in the first-step-transfer DNA of bacteriophage T5. J Mol Biol. 1969 Aug 28;44(1):173–183. doi: 10.1016/0022-2836(69)90412-4. [DOI] [PubMed] [Google Scholar]
  9. Radding C. M. Regulation of lambda exonuclease. I. Properties of lambda exonuclease purified from lysogens of lambda T11 and wild type. J Mol Biol. 1966 Jul;18(2):235–250. doi: 10.1016/s0022-2836(66)80243-7. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. 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]
  13. 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]
  14. 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]
  15. 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]
  16. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  17. THOMAS C. A., Jr, RUBENSTEIN I. THE ARRANGEMENTS OF NUCLEOTIDE SEQUENCES IN T2 AND T5 BACTERIOPHAGE DNA MOLECULES. Biophys J. 1964 Mar;4:93–106. doi: 10.1016/s0006-3495(64)86771-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wilson G. A., Young F. E. Isolation of a sequence-specific endonuclease (BamI) from Bacillus amyloliquefaciens H. J Mol Biol. 1975 Sep 5;97(1):123–125. doi: 10.1016/s0022-2836(75)80028-3. [DOI] [PubMed] [Google Scholar]
  19. 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]

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