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Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNA
Open accessAbstract
DNA fragments obtained from Eco RI endonuclease digestion of bacteriophage ϕ80pt190 ( trp + ) and the plasmid ColE1 were covalently joined with polynucleotide ligase. Transformation of Escherichia coli trp - strains to tryptophan independence with the recombined DNA selected for reconstituted ColE1 plasmids containing the tryptophan operon and the ϕ80 immunity region. Similarly, an Eco RI endonuclease generated fragment of plasmid pSC105 DNA containing the genetic determinant of kanamycin resistance was inserted into the ColE1 plasmid and recovered in E. coli . The plasmids containing the trp operon (ColE1- trp ) and the kanamycin resistance gene were maintained under logarithmic growth conditions at a level of 25-30 copies per cell and accumulate to the extent of several hundred copies per cell in the presence of chloramphenicol. Cells carrying the ColE1- trp plasmid determined the production of highly elevated levels of trp operon-specific mRNA and tryptophan biosynthetic enzymes.
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Citations
Cited by
Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene.
Grunstein, M., & Hogness, D. S. (1975) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.72.10.3961 · Cited by 3,431
Expression in Escherichia coli of chemically synthesized genes for human insulin.
Goeddel, D. V., Kleid, D. G., Bolivar, F., Heyneker, H. L., Yansura, D. G., Crea, R., Hirose, T., Kraszewski, A., Itakura, K., & Riggs, A. D. (1979) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.76.1.106 · Cited by 869
Engineering BioBrick vectors from BioBrick parts
Shetty, R. P., Endy, D., & Knight, T. F. (2008) · Journal of Biological Engineering · doi:10.1186/1754-1611-2-5 · Cited by 760
FLENSBURG, J., & SKÖLD, O. (1987) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1987.tb10664.x · Cited by 103
Generating functional plasmid origins with OriGen
Irvine, J., Martinson, J. N. V., Arora, J., Hasham, S. I., Patel, J. R., Qi, E. T., Cress, B. F., & Rubin, B. E. (2025) · Nucleic Acids Research · doi:10.1093/nar/gkaf1198 · Cited by 4
Metadata
- Authors
- Hershfield, V., Boyer, H. W., Yanofsky, C., Lovett, M. A., & Helinski, D. R.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1974-09
- Volume
- 71
- Issue
- 9
- Pages
- 3455-3459
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1994503800
- Cited by
- 540
- References
- 0
- FWCI
- 39.24
- Type
- journal article
- Language
- en
- Open access
- green
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- Hershfield, V., Boyer, H. W., Yanofsky, C., Lovett, M. A., & Helinski, D. R. (1974). Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNA. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.71.9.3455
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