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. 1978 Feb;75(2):774–778. doi: 10.1073/pnas.75.2.774

Construction and characterization of an Escherichia coli plasmid bearing a functional gene G of bacteriophage phiX174.

M Z Humayun, R W Chambers
PMCID: PMC411339  PMID: 273240

Abstract

In order to study the mutagenic effects of site-specific, covalent modifications of biologically active DNA, we need host cells that are permissive for any type of mutation that might be produced in vivo from the modified DNA. Specifically, we require a general, in vivo complementation system for the bacteriophage phiX174 gene G, an essential gene that we have chosen for our initial studies of chemical mutagenesis. Toward this end, we have constructed a plasmid (pphiXG) that carries a functional copy of phiX174 gene G. Three different bacterial strains that are nonpermissive for am9, a gene G amber mutant, have been transformed with pphiXG. The transformants are now permissive for this gene G mutant, but not for the gene A or E mutants that have been tested. This paper describes the construction and the biochemical characterization of this plasmid, pphiXG, and describes some of the biological properties exhibited by the pphiXG-bearing strains.

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Selected References

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

  1. Axelrod N. Transcription of bacteriophage phiX174 in vitro: analysis with restriction enzymes. J Mol Biol. 1976 Dec 25;108(4):771–779. doi: 10.1016/s0022-2836(76)80116-7. [DOI] [PubMed] [Google Scholar]
  2. BOLLUM F. J. Thermal conversion of nonpriming deoxyribonucleic acid to primer. J Biol Chem. 1959 Oct;234:2733–2734. [PubMed] [Google Scholar]
  3. Baas P. D., van Heusden G. P., Vereijken J. M., Weisbeek P. J., Jansz H. S. Cleavage map of bacteriophage phiX174 RF DNA by restriction enzymes. Nucleic Acids Res. 1976 Aug;3(8):1947–1960. doi: 10.1093/nar/3.8.1947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benbow R. M., Hutchison C. A., Fabricant J. D., Sinsheimer R. L. Genetic Map of Bacteriophage phiX174. J Virol. 1971 May;7(5):549–558. doi: 10.1128/jvi.7.5.549-558.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Benbow R. M., Mayol R. F., Picchi J. C., Sinsheimer R. L. Direction of Translation and Size of Bacteriophage phiX174 Cistrons. J Virol. 1972 Jul;10(1):99–114. doi: 10.1128/jvi.10.1.99-114.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benbow R. M., Zuccarelli A. J., Davis G. C., Sinsheimer R. L. Genetic recombination in bacteriophage phi chi 174. J Virol. 1974 Apr;13(4):898–907. doi: 10.1128/jvi.13.4.898-907.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Denhardt D. T. The single-stranded DNA phages. CRC Crit Rev Microbiol. 1975 Dec;4(2):161–223. doi: 10.3109/10408417509111575. [DOI] [PubMed] [Google Scholar]
  10. Godson G. N., Boyer H. Susceptibility of the phiX-like phages G4 and G14 to R-EcoRi endonuclease. Virology. 1974 Nov;62(1):270–275. doi: 10.1016/0042-6822(74)90321-3. [DOI] [PubMed] [Google Scholar]
  11. Higuchi R., Paddock G. V., Wall R., Salser W. A general method for cloning eukaryotic structural gene sequences. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3146–3150. doi: 10.1073/pnas.73.9.3146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Humayun Z., Jeffrey A., Ptashne M. Completed DNA sequences and organization of repressor-binding sites in the operators of phage lambda. J Mol Biol. 1977 May 15;112(2):265–277. doi: 10.1016/s0022-2836(77)80143-5. [DOI] [PubMed] [Google Scholar]
  13. Jackson D. A., Symons R. H., Berg P. Biochemical method for inserting new genetic information into DNA of Simian Virus 40: circular SV40 DNA molecules containing lambda phage genes and the galactose operon of Escherichia coli. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2904–2909. doi: 10.1073/pnas.69.10.2904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lee A. S., Sinsheimer R. L. A cleavage map of bacteriophage phiX174 genome. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2882–2886. doi: 10.1073/pnas.71.7.2882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Little J. W., Lehman I. R., Kaiser A. D. An exonuclease induced by bacteriophage lambda. I. Preparation of the crystalline enzyme. J Biol Chem. 1967 Feb 25;242(4):672–678. [PubMed] [Google Scholar]
  16. Maniatis T., Kee S. G., Efstratiadis A., Kafatos F. C. Amplification and characterization of a beta-globin gene synthesized in vitro. Cell. 1976 Jun;8(2):163–182. doi: 10.1016/0092-8674(76)90001-5. [DOI] [PubMed] [Google Scholar]
  17. McCann J., Choi E., Yamasaki E., Ames B. N. Detection of carcinogens as mutagens in the Salmonella/microsome test: assay of 300 chemicals. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5135–5139. doi: 10.1073/pnas.72.12.5135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Roychoudhury R., Jay E., Wu R. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acids Res. 1976 Jan;3(1):101–116. doi: 10.1093/nar/3.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sanger F., Air G. M., Barrell B. G., Brown N. L., Coulson A. R., Fiddes C. A., Hutchison C. A., Slocombe P. M., Smith M. Nucleotide sequence of bacteriophage phi X174 DNA. Nature. 1977 Feb 24;265(5596):687–695. doi: 10.1038/265687a0. [DOI] [PubMed] [Google Scholar]
  20. Smith L. H., Sinsheimer R. L. The in vitro transcription units of bacteriophage phiX174. II. In vitro initiation sites of phiX174 transcription. J Mol Biol. 1976 Jun 5;103(4):699–710. doi: 10.1016/0022-2836(76)90204-7. [DOI] [PubMed] [Google Scholar]
  21. Vosberg H. P., Hoffmann-Berling H. DNA synthesis in nucleotide-permeable Escherichia coli cells. I. Preparation and properties of ether-treated cells. J Mol Biol. 1971 Jun 28;58(3):739–753. doi: 10.1016/0022-2836(71)90037-4. [DOI] [PubMed] [Google Scholar]
  22. Wensink P. C., Finnegan D. J., Donelson J. E., Hogness D. S. A system for mapping DNA sequences in the chromosomes of Drosophila melanogaster. Cell. 1974 Dec;3(4):315–325. doi: 10.1016/0092-8674(74)90045-2. [DOI] [PubMed] [Google Scholar]
  23. Witkin E. M. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. doi: 10.1128/br.40.4.869-907.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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