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High-voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA.
Open accessAbstract
Electroporation permits the uptake of DNA by mammalian cells and plant protoplasts because it induces transient permeability of the cell membrane. We investigated the utility of high-voltage electroporation as a method for genetic transformation of intact bacterial cells by using the enteric pathogen Campylobacter jejuni as a model system. This report demonstrates that the application of high-voltage discharges to bacterial cells permits genetic transformation. Our method involves exposure of a Campylobacter cell suspension to a high-voltage exponential decay discharge (5-13 kV/cm) for a brief period of time (resistance-capacitance time constant = 2.4-26 msec) in the presence of plasmid DNA. Electrical transformation of C. jejuni results in frequencies as high as 1.2 x 10(6) transformants per microgram of DNA. We have investigated the effects of pulse amplitude and duration, cell growth conditions, divalent cations, and DNA concentration on the efficiency of transformation. Transformants of C. jejuni obtained by electroporation contained structurally intact plasmid molecules. In addition, evidence is presented that indicates that C. jejuni possesses DNA restriction and modification systems. The use of electroporation as a method for transforming other bacterial species and guidelines for its implementation are also discussed.
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Citations
Cited by
Pathogenesis of Enteric Infection by Campylobacter
Ketley, J. M. (1997) · Microbiology · doi:10.1099/00221287-143-1-5 · Cited by 411
Wassenaar, T. M., Bleumink-Pluym, N. M., & van der Zeijst, B. A. (1991) · The EMBO Journal · doi:10.1002/j.1460-2075.1991.tb07736.x · Cited by 296
Cell poration and cell fusion using an oscillating electric field
Chang, D. C. (1989) · Biophysical Journal · doi:10.1016/s0006-3495(89)82711-0 · Cited by 214
Introduction of plasmids into whole cells of Clostridium acetobutylicum by electroporation
Oultram, J. D., Loughlin, M., Swinfield, T.-J., Brehm, J. K., Thompson, D. E., & Minton, N. P. (1988) · FEMS Microbiology Letters · doi:10.1111/j.1574-6968.1988.tb03154.x · Cited by 133
Metadata
- Authors
- Miller, J. F., Dower, W. J., & Tompkins, L. S.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1988-02
- Volume
- 85
- Issue
- 3
- Pages
- 856-860
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1974903923
- Cited by
- 214
- References
- 0
- FWCI
- 22.04
- Type
- journal article
- Language
- en
- Open access
- green
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- CoChat. (2026). Citation check: High-voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA. https://app.cochat.ai/papers/10.1073/pnas.85.3.856
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- Miller, J. F., Dower, W. J., & Tompkins, L. S. (1988). High-voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.85.3.856
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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