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Construction and some properties of packageable plasmid F

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Summary

A derivative of plasmid F which is packageable in λ phage coat was constructed using techniques of in vitro recombination. This plasmid is composed of three DNA fragments generated by restriction enzyme EcoRI: a miniF fragment (fragment f5 of F'lac) which is able to replicate autonomously, a DNA fragment from Staphylococcus Plasmid that carries the β-lactamase gene, and a portion of guaA (B) transducing λ phage DNA carrying λ cohesive ends (cos site) along with almost all the late genes but devoid of all those genes and sites that are needed for replication, regulation, and recombination. The hybrid plasmid has a molecular weight of 2.7×107 daltons, about 84% size of λ phage genome, and can be packaged in λ coat when helper phage replicates in the plasmid-carrier cell. The package plasmid and the helper λ phage particles are separated by CsCl density gradient centrifugation. The replication characteristics of the recombinant plasmid are all those of F including the copy number, incompatibility, and curing with acidine orange. The packaged plasmid is injected into an F- cell and establishes a plasmid state with normal efficiency. In F+ or Hfr cells, the resident F factor hinders this process.

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References

  • Cohen, S.N., Chang, A.C.Y., Hsu, L.: Nonchromosomal antibiotic resistance in bacteria: Genetic transformation of Escherichia coli by R-factor DNA. Proc. Natl. Acad. Sci. U.S.A. 69, 2110–2114 (1972)

    Google Scholar 

  • Davidson, N., Szybalski W.: Physical and chemical characteristics of lambda DNA In: The bacteriophage lambda (Hershey, A.D., ed.), pp. 45–82. New York: Cold Spring Harbor Laboratory 1971

    Google Scholar 

  • Feiss, M., Fischer, R.A., Crayton, M.A., Eigner, C.: Packaging of the λ chromosome: Effect of chromosome length. Virology 77, 281–293 (1977)

    Google Scholar 

  • Fukumaki, Y., Shimada, K., Takagi, Y.: Specialized transduction of colicine El DNA in Escherichia coli K12 by phage Lambda. Proc. Natl. Acad. Sci. U.S.A. 73, 3238–3242 (1976)

    Google Scholar 

  • Greene, P.J., Bethlach, M.C., Goodman, H.M., Boyer, H.W.: The EcoRI restriction endonuclease. In: DNA replication (Wickner, R.B. ed.), pp. 87–103. New York: Marcel Dekker, Inc. 1974

    Google Scholar 

  • Guyer, M.S., Figurski, K., Davidson, N.: Electron microscope study of a plasmid chimera containing the replication region of the Escherichia coli F plasmid. J. Bacteriol. 127, 988–997 (1976)

    Google Scholar 

  • Haggerty, D.M., Schlief, R.B.: Location in bacteriophage Lambda DNA of cleavage site of the site-specific endonucleasc from Bacillus amyloliquifaciens HI. J. Virol. 18, 659–663 (1976)

    Google Scholar 

  • Manis, J.J., Kline, B.C.: Restriction endonuclease mapping and mutagenesis of the F sex factor replication region. Mol. Gen. Genet. 152, 175–182 (1977)

    Google Scholar 

  • Matsubara, K., Takagi, Y., Mukai, T.: In vitro construction of different oligomeric forms of λ dv DNA and studies on their transforming activities. J. Virol. 16, 479–485 (1975)

    Google Scholar 

  • Mukai, T., Matsubara, K., Takagi, Y.: In vitro construction of ColEl factor carrying genes for synthesis of guanine or thymine. Proc. Jap. Acad. 51, 353–357 (1975)

    Google Scholar 

  • Mukai, T., Matsubara, K., Takagi, Y.: Cloning bacterial genes with plasmid λdv. Mol. Gen. Genet. 146, 269–274 (1976)

    Google Scholar 

  • Robinson, L.H., Landy, A.: HindII, HindIII and Hpal restriction fragment maps of bacteriophage Lambda DNA. Gene 2, 1–31 (1977)

    Google Scholar 

  • Sharp, P.A., Sugden, B., Sambrook, J.: Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose-ethidium bromide electrophoresis. Biochemistry 12, 3055–3063 (1973)

    Google Scholar 

  • Timmis, K., Cabello, F., Cohen, S.N.: Cloning, isolation and characterization of replication regions of complex plasmid genomes. Proc. Natl. Acad. Sci. U.S.A. 72, 2242–2246 (1975)

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Molecular Genetics, Osaka University Medical School, Kita-ku, 530, Osaka, Japan

    Yasuhiko Hayakawa & Kenichi Matsubara

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  1. Yasuhiko Hayakawa
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  2. Kenichi Matsubara
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Communicated by T. Yura

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Hayakawa, Y., Matsubara, K. Construction and some properties of packageable plasmid F. Molec. Gen. Genet. 169, 107–112 (1979). https://doi.org/10.1007/BF00267551

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  • DOI: https://doi.org/10.1007/BF00267551

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