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. 1981 Dec;40(3):839–847. doi: 10.1128/jvi.40.3.839-847.1981

Genetic recombination of bacteriophage T7 in vivo studied by use of a simple physical assay.

D Lee, P D Sadowski
PMCID: PMC256695  PMID: 6275119

Abstract

A new physical method was developed to assay genetic recombination of phage T7 in vivo. The assay utilized T7 mutants that carry unique restriction sites and was based on the detection of a new restriction fragment generated by recombination. Using this assay, we reexamined the genetic requirements for recombination of T7 DNA. Our results were in total agreement with previous findings in that recombination required the products of genes 3 (endonuclease), 4 (primase), 5 (DNA polymerase), and 6 (exonuclease). Recombination was found to be independent of DNA ligase and DNA packaging and maturation functions.

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Selected References

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  1. Berger H., Warren A. J., Fry K. E. Variations in genetic recombination due to amber mutations in T4D bacteriophage. J Virol. 1969 Feb;3(2):171–175. doi: 10.1128/jvi.3.2.171-175.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bernstein H. Repair and recombination in phage T4. I. Genes affecting recombination. Cold Spring Harb Symp Quant Biol. 1968;33:325–331. doi: 10.1101/sqb.1968.033.01.037. [DOI] [PubMed] [Google Scholar]
  3. Bingham A. H., Sharman A. F., Atkinson T. The purification of restriction endonuclease EcoRI by precipitation involving polyethyleneimine. FEBS Lett. 1977 Apr 15;76(2):250–256. doi: 10.1016/0014-5793(77)80162-2. [DOI] [PubMed] [Google Scholar]
  4. Broker T. R. An electron microscopic analysis of pathways for bacteriophage T4 DNA recombination. J Mol Biol. 1973 Nov 25;81(1):1–16. doi: 10.1016/0022-2836(73)90243-x. [DOI] [PubMed] [Google Scholar]
  5. Burck K. B., Miller R. C., Jr Marker rescue and partial replication of bacteriophage T7 DNA. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6144–6148. doi: 10.1073/pnas.75.12.6144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hausmann R., LaRue K. Variations in sedimentation patterns among deoxyribonucleic acids synthesized after infection of Escherichia coli by different amber mutants of bacteriophage T7. J Virol. 1969 Feb;3(2):278–281. doi: 10.1128/jvi.3.2.278-281.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jeffreys A. J., Wilson V., Wood D., Simons J. P., Kay R. M., Williams J. G. Linkage of adult alpha- and beta-globin genes in X. laevis and gene duplication by tetraploidization. Cell. 1980 Sep;21(2):555–564. doi: 10.1016/0092-8674(80)90493-6. [DOI] [PubMed] [Google Scholar]
  8. Kerr C., Sadowski P. D. The involvement of genes 3,4,5 and 6 in genetic recombination in bacteriophage T7. Virology. 1975 May;65(1):281–285. doi: 10.1016/0042-6822(75)90031-8. [DOI] [PubMed] [Google Scholar]
  9. Kikuchi Y., Nash H. Integrative recombination of bacteriophage lambda: requirement for supertwisted DNA in vivo and characterization of int. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1099–1109. doi: 10.1101/sqb.1979.043.01.122. [DOI] [PubMed] [Google Scholar]
  10. Langman L., Paetkau V., Scraba D., Miller R. C., Jr, Roeder G. S., Sadowski P. D. The structure and maturation of intermediates in bacteriophage T7 DNA replication. Can J Biochem. 1978 Jun;56(6):508–516. doi: 10.1139/o78-078. [DOI] [PubMed] [Google Scholar]
  11. Masamune Y., Frenkel G. D., Richardson C. C. A mutant of bacteriophage T7 deficient in polynucleotide ligase. J Biol Chem. 1971 Nov 25;246(22):6874–6879. [PubMed] [Google Scholar]
  12. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  13. Mizuuchi K., Nash H. A. Restriction assay for integrative recombination of bacteriophage lambda DNA in vitro: requirement for closed circular DNA substrate. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3524–3528. doi: 10.1073/pnas.73.10.3524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Paetkau V., Langman L., Bradley R., Scraba D., Miller R. C., Jr Folded, concatenated genomes as replication intermediates of bacteriophage T7 DNA. J Virol. 1977 Apr;22(1):130–141. doi: 10.1128/jvi.22.1.130-141.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Powling A., Knippers R. Some functions involved in bacteriophage T7 genetic recombination. Mol Gen Genet. 1974;134(2):173–180. doi: 10.1007/BF00268418. [DOI] [PubMed] [Google Scholar]
  16. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  17. Roberts L., Sheldon R., Sadowski P. D. Genetic recombination of bacteriophage T7 DNA in vitro. IV. Asymmetry of recombination frequencies caused by polarity of DNA packaging. Virology. 1978 Aug;89(1):252–261. doi: 10.1016/0042-6822(78)90057-0. [DOI] [PubMed] [Google Scholar]
  18. Roeder G. S., Sadowski P. D. Bacteriophage T7 morphogenesis: phage-related particles in cells infected with wild-type and mutant T7 phage. Virology. 1977 Jan;76(1):263–285. doi: 10.1016/0042-6822(77)90302-6. [DOI] [PubMed] [Google Scholar]
  19. Roeder G. S., Sadowski P. D. Pathways of recombination of bacteriophage T7 DNA in vitro. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1023–1032. doi: 10.1101/sqb.1979.043.01.111. [DOI] [PubMed] [Google Scholar]
  20. Rosenberg A. H., Simon M. N., Studier F. W., Roberts R. J. Survey and mapping of restriction endonuclease cleavage sites in bacteriophage T7 DNA. J Mol Biol. 1979 Dec 25;135(4):907–915. doi: 10.1016/0022-2836(79)90519-9. [DOI] [PubMed] [Google Scholar]
  21. Sadowski P. D., Kerr C. Degradation of Escherichia coli B deoxyribonucleic acid after infection with deoxyribonucleic acid-defective amber mutants of bacteriophage T7. J Virol. 1970 Aug;6(2):149–155. doi: 10.1128/jvi.6.2.149-155.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sadowski P. D., Vetter D. Genetic recombination of bacteriophage T7 DNA in vitro. Proc Natl Acad Sci U S A. 1976 Mar;73(3):692–696. doi: 10.1073/pnas.73.3.692. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Serwer P. Fast sedimenting bacteriophage T7 DNA from T7-infected Escherichia coli. Virology. 1974 May;59(1):70–88. doi: 10.1016/0042-6822(74)90207-4. [DOI] [PubMed] [Google Scholar]
  24. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  25. Stahl F. W. Special sites in generalized recombination. Annu Rev Genet. 1979;13:7–24. doi: 10.1146/annurev.ge.13.120179.000255. [DOI] [PubMed] [Google Scholar]
  26. Studier F. W. Genetic analysis of non-essential bacteriophage T7 genes. J Mol Biol. 1973 Sep 15;79(2):227–236. doi: 10.1016/0022-2836(73)90002-8. [DOI] [PubMed] [Google Scholar]
  27. Studier F. W. The genetics and physiology of bacteriophage T7. Virology. 1969 Nov;39(3):562–574. doi: 10.1016/0042-6822(69)90104-4. [DOI] [PubMed] [Google Scholar]
  28. TOMIZAWA J. I., ANRAKU N. MOLECULAR MECHANISMS OF GENETIC RECOMBINATION IN BACTERIOPHAGE. II. JOINING OF PARENTAL DNA MOLECULES OF PHAGE T4. J Mol Biol. 1964 Apr;8:516–540. doi: 10.1016/s0022-2836(64)80009-7. [DOI] [PubMed] [Google Scholar]
  29. Tsujimoto Y., Ogawa H. EcoRI-sensitive mutation of T7 phage. Mol Gen Genet. 1977 Jan 18;150(2):221–223. doi: 10.1007/BF00695402. [DOI] [PubMed] [Google Scholar]
  30. Tsujimoto Y., Ogawa H. Intermediates in genetic recombination of bacteriophage T7 DNA. J Mol Biol. 1977 Jan 25;109(3):423–426. doi: 10.1016/s0022-2836(77)80021-1. [DOI] [PubMed] [Google Scholar]

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