Transformation of Escherichia coli by Recombinant Plasmid Replicons Constructed in Vitro

  • Stanley N. Cohen


Treatment of Escherichia coli with calcium chloride renders this bacterial species competent for transfection by purified bacteriophage DNA (Mandel and Higa, 1970) and for transformation by plasmid (Cohen et al., 1972) or E. coli chromosomal (Cosloy and Oishi, 1973; Wackernagel, 1973) DNA. Transformation of E. coli by plasmid DNA, which unlike chromosomal DNA does not require a recipient strain defective in the recBC nuclease, has proved to be a useful tool for investigating the genetic and molecular properties of discrete plasmid species, and for introducing nonconjugative Plasmids into specific E. coli hosts in the absence of transducing phage or conjugally proficient transfer plasmids (Cohen et al., 1972; Cohen and Chang, 1973; van Embden and Cohen, 1973; Lederberg et al., 1973; Guerry et al., 1974; Oishi and Cosloy, 1974).


Buoyant Density EcoRI Restriction Endonuclease Resistance Plasmid EcoRI Endonuclease Plasmid Replicon 
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  1. Birnstiel, M. L., M. Chipchase and J. Speirs. 1971. The ribosomal RNA cistrons. Prog. Nucl. Res. Mol. Biol. 11: 351.CrossRefGoogle Scholar
  2. Chang, A. C. Y. and S. N. Cohen. 1974. Genome construction between bacterial species in vitro: replication and expression of Staphylococcus plasmid genes in E. coli. Proc. Nat. Acad. Sci. U.S.A. 71 in press.Google Scholar
  3. Cohen, S. N. and A. C. Y. Chang. 1973. Recircularization and autonomous replication of a sheared R-factor DNA segment in Escherichia coli transformants. Proc. Nat. Acad. Sci. U.S.A. 70: 1293.CrossRefGoogle Scholar
  4. Cohen, S. N., A. C. Y. Chang, H. W. Boyer, and R. B. Helling. 1973. Construction of biologically functional bacterial plasmids in vitro. Proc. Nat. Acad. Sci. U.S.A. 70: 3240.CrossRefGoogle Scholar
  5. Cohen, S. N., A. C. Y. Chang and L. Hsu. 1972. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc. Nat. Acad. Sci. U.S.A. 69: 2110.CrossRefGoogle Scholar
  6. Cosloy, S. D. and M. Oishi. 1973. Genetic transformation in Escherichia coli K12. Proc. Nat. Acad. Sci. U.S.A. 70: 84.CrossRefGoogle Scholar
  7. Dawid, I. B., D. D. Brown and R. H. Reeder. 1970. Composition and structure of chromosomal and amplified ribosomal DNA’s of Xenopus laevis. J. Mol. Biol. 51: 341.PubMedCrossRefGoogle Scholar
  8. Guerry, P., J. van Embden and S. Falkow. 1974. Molecular nature of two nonconjugative plasmids carrying drug resistance genes. J. Bacteriol. 117: 619.PubMedGoogle Scholar
  9. Hedgepeth, J., H. M. Goodman and H. W. Boyer. 1972. DNA nucleotide sequence restricted by the RI endonuclease. Proc. Nat. Acad. Sci. U.S.A. 69: 3448.CrossRefGoogle Scholar
  10. Hourcade, D., D. Dressier and J. Wolfson. 1973. The amplification of ribosomal RNA genes involves a rolling circle intermediate. Proc. Nat. Acad. Sci. U.S.A. 70: 2926.CrossRefGoogle Scholar
  11. Lederberg, E. M., L. L. Brothers and S. N. Cohen. 1973. Molecular properties of an F-lac+-tetracycline resistance plasmid in E. coli and Salmonella typhimurium LT2. Genetics 74: 5152 (abstract).Google Scholar
  12. Lindberg, M. and R. P. Novick. 1973. Plasmid-specific transformation in Staphylococcus aureus. J. Bacteriol. 115: 139.PubMedGoogle Scholar
  13. Mandel, M. and A. Higa. 1970. Calcium-dependent bacteriophage DNA infection. J. Mol. Biol. 53: 159.PubMedCrossRefGoogle Scholar
  14. Mertz, J. E. and R. W. Davis. 1972. Cleavage of DNA by RI restriction endonuclease generates cohesive ends. Proc. Nat. Acad. Sci. U.S.A. 69: 3370.CrossRefGoogle Scholar
  15. Morrow, J. F., S. N. Cohen, A. C. Y. Chang, H. W. Boyer, H. M. Goodman and R. B. Helling. 1974. Replication and transcription of eukaryotic DNA in Escherichia coli. Proc. Nat. Acad. Sci. U.S.A. 71 in press.Google Scholar
  16. Novick, R. P. and D. Bouanchaud. 1971. Extrachromosomal nature of drug resistance in Staphylococcus aureus. Ann. N. Y. Acad. Sci. 182: 279.PubMedCrossRefGoogle Scholar
  17. Oishi, M. and S. D. Cosloy. 1974. Specialized transformation in E. coli K12. Nature 248: 112.PubMedCrossRefGoogle Scholar
  18. Rush, M. G., C. N. Gordon, R. N. Novick and R. C. Warner. 1969. Penicillinase plasmid DNA from Staphylo coccus aureus. Proc. Nat. Acad. Sci. U.S.A. 63: 1304.CrossRefGoogle Scholar
  19. Sgaramella, V. 1972. Enzymatic oligomerization of bacteriophage P22 DNA and of linear simian virus 40 DNA. Proc. Nat. Acad. Sci. U.S.A. 69: 3389.CrossRefGoogle Scholar
  20. Sharp, P. A., S. N. Cohen and N. Davidson. 1973. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. II. Structure of drug resistance (R) factors and F factors. J. Mol. Biol. 75: 235.PubMedCrossRefGoogle Scholar
  21. Silver, R. P. and S. N. Cohen. 1972. Nonchromosomal antibiotic resistance in bacteria. V. Isolation and characterization of R-factor mutants exhibiting temperature-sensitive repression of fertility. J. Bacteriol. 110: 1082.PubMedGoogle Scholar
  22. van Embden, J. and S. N. Cohen. 1973. Molecular and genetic structures of an R factor system consisting of independent transfer and drug resistance plasmids. J. Bacteriol. 116: 699.PubMedGoogle Scholar
  23. Wackernagel, W. 1973. Genetic transformation in E. coli: the inhibitory role of the recBC DNAase. Biochem. Biophys. Res. Commun. 51: 306.PubMedCrossRefGoogle Scholar
  24. Wensink, P. C. and D. D. Brown. 1971. Denaturation map of the ribosomal DNA of Xenopus laevis. J. Mol. Biol. 60: 235.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1974

Authors and Affiliations

  • Stanley N. Cohen
    • 1
  1. 1.Department of MedicineStanford University School of MedicineStanfordUSA

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