Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

High Efficiency Transformation of Intact Yeast Cells by Electric Field Pulses

Bio/Technology volume 8, pages 223–227 (1990)Cite this article

Abstract

We have developed an efficient method that electrically introduces DNA into intact yeast cells. Saccharomyces cerevisiae was used as a model in order to optimize the transformation protocol. Transformation efficiencies of 107 transformants/μg of plasmid DNA were obtained with a square wave electric pulse of 2.7 kV/cm during 15 milliseconds. The technique is simple and rapid. Even small quantities of DNA (100 pg) can be used to transform 108 cells. Important parameters are the pulse field strength and duration. Pretreatment of the yeast cells in the early phase of exponential growth with dithiothreitol increases transformation efficiency. The method has been successfully applied to various strains of S. cerevisiae as well as to other types of yeast.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Ito, H., Jukuda, A., Murata, K. and Kimura, A. 1983. Transformation of intact yeast cells treated with alkali cations. J. Bact. 153: 163–168.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Beggs, J.D. 1978. Transformation of yeast by a replicating hybrid plasmid. Nature 275: 104–109.

    Article  CAS  PubMed  Google Scholar 

  3. Hinnen, A., Hicks, J.B. and Fink, G.R. 1978. Transformation of yeast. Proc. Natl. Acad. Sci. USA 75: 1929–1933.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Costanzo, M.C. and Fox, T.D. 1988. Transformation of yeast by agitation with glass beads. Genetics 120: 667–670.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Neumann, E., Schaefer-Ridder, M., Wang, Y. and Hofschneider, P.H. 1982. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1: 841–845.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Venkat-Gopal, T. 1985. Gene transfer method for transient gene expression, stable transformation and co transformation of suspension cell cultures. Mol. Cell. Biol. 5: 1188–1190.

    Article  Google Scholar 

  7. Zerbib, D., Amalric, F. and Teissié, J. 1985. Electric field mediated transformation: isolation and characterization of a TK+ subclone. Biochem. Biophys. Res. Comm. 129: 611–618.

    Article  CAS  PubMed  Google Scholar 

  8. Narayanan, R., Jastreboff, M.M., Chiu, C.F. and Bertino, J.R. 1986. In vivo expression of a nonselected gene transferred into murine hematopoietic stem cells by electroporation. Biochem. Biophys. Res. Comm. 141: 1018–1024.

    Article  CAS  PubMed  Google Scholar 

  9. Toneguzzo, F. and Keating, A. 1986. Stable expression of selectable genes introduced into human hematopoietic stem cells by electric field mediated DNA transfer. Proc. Natl. Acad. Sci. USA 83: 3496–3499.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Toneguzzo, F., Hayday, A.C. and Keating, A. 1986. Electric field mediated DNA transfer: transient and stable expression in human and mouse lymphoid cells. Mol. Cell. Biol. 6: 703–706.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Tur-kaspa, R., Teicher, L., Levine, B.J., Skoultchi, A.I. and Shafritz, D.A. 1986. Use of electroporation to introduce biologically active foreign genes into primary rat hepatocytes. Mol. Cell. Biol. 6: 716–718.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Chu, G., Hayakawa, H. and Berg, P. 1987. Electroporation for the efficient transfection of mammalian cells with DNA. Nucl. Acid. Res. 15: 1311–1326.

    Article  CAS  Google Scholar 

  13. Spandidos, D.A. 1987. Electric-field mediated gene transfer (electroporation) into mouse Friend and human K 562 erythroleukemic cells. Gene Anal. Tech. 4: 50–56.

    Article  CAS  PubMed  Google Scholar 

  14. Schocher, R.J., Shillito, R.D., Saul, M.W., Paszkowski, J. and Potrykus, I. 1986. Co-transformation of unliked foreign genes into plants by direct gene transfer. Bio/Technology 4: 1093–1096.

    CAS  Google Scholar 

  15. Fromm, M.E., Taylor, L.P. and Walbot, V. 1986. Stable transformation of maize after gene transfer by electroporation. Nature 319: 791–793.

    Article  CAS  PubMed  Google Scholar 

  16. Riggs, C.D. and Bates, G.W. 1986. Stable transformation of tobacco by electroporation: evidence for plasmid concatenation. Proc. Natl. Acad. Sci. USA 83: 5602–5606.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Toriyama, K., Arimoto, Y., Uchimiya, H. and Hinata, K. 1988. Transgenic rice plants after direct gene transfer into protoplasts. Bio/Technology 6: 1072–1074.

    CAS  Google Scholar 

  18. Calvin, M.N. and Hanawalt, P.C. 1988. High-Efficiency transformation of bacterial cells by electroporation. J. Bact. 170: 2796–2801.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Friedler, S. and Wirth, R. 1988. Transformation of bacteria with plasmid DNA by electroporation. Anal. Biochem. 170: 38–44.

    Article  Google Scholar 

  20. Miller, J.F., Dower, W.J. and Tompkins, L.S. 1988. High voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA. Proc. Natl. Acad. Sci. USA 85: 856–860.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Powell, I.B., Achen, M.G., Hillier, A.J. and Davidson, B.E. 1988. A simple and rapid method for genetic transformation of lactic streptococci by electroporation. Applied and Env. Microbiol. 54: 655–660.

    CAS  Google Scholar 

  22. Gibson, W.C., White, T.C., Laird, P.W. and Borst, P. 1987. Stable introduction of exogenous DNA into Trypanosoma brucei . EMBO J. 6: 2457–2461.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Forster, W. and Neumann, E. 1989. Gene transfer by electroporation, p. 299–318. In: Electroporation and Electrofusion in Cell Biology. Neumann, E., Sowers, A.E. and Jordan, C.A. (Eds.). Plenum Press, NY.

    Chapter  Google Scholar 

  24. Chassy, B.M., Mercenier, A. and Flickinger, J. 1988. Transformation of bacteria by electroporation. Trends in Biotech. 6: 303–309.

    Article  CAS  Google Scholar 

  25. Andreason, G.L. and Evans, G.A. 1988. Introduction and expression of DNA molecules in eukaryotic cells by electroporation. Biotechniques 6: 650–660.

    CAS  PubMed  Google Scholar 

  26. Karube, I., Tamiya, E. and Matsuoka, H. 1985. Transformation of Saccharomyces cerevisiae spheroplasts by high electric pulse. FEBS Letts. 182: 90–94.

    Article  CAS  Google Scholar 

  27. Hashimoto, H., Morijawa, H., Yamata, Y. and Kimura, A. 1985. A novel method for transformation of intact yeast cells by electroinjection of plasmid DNA. Applied Microbiol. Biotechnol. 2: 336–339.

    Google Scholar 

  28. Weaver, J.C., Harrison, G.I., Bliss, J.G., Mourant, J.R. and Powell, K.T. 1988. Electroporation: high frequency of occurrence of a transient high permeability state in erythrocytes and intact yeast. FEBS Letts. 1: 30–34.

    Article  Google Scholar 

  29. Boyer, H.W. and Roulland-Dussoix, D. 1969. A complementation analysis of the restriction and modification of DNA in Escherichia coli . J. Mol. Biol. 41: 459–472.

    Article  CAS  PubMed  Google Scholar 

  30. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

    Google Scholar 

  31. Sherman, F., Fink, G.R. and Hicks, J.B. 1986. Methods in Yeast Genetics (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).

    Google Scholar 

  32. Teissié, J., Rols, M.P. 1988. Electropermeabilization and electrofusion of cells, p. 251–268. In: Dynamics of Membranes Proteins and Cellular Energetics. Latruffe, N., Gaudemer, Y., Vignais, P., Azzi, A. (Eds.). Springer Verlag, Berlin.

    Chapter  Google Scholar 

  33. EBC. Analytica Microbiologica. J. Inst. Brew. 1987. 83: 109–118.

  34. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Strulh, K. 1989. Protocols in Molecular Biology. Vol 2. John Wiley and Sons, NY.

    Google Scholar 

Download references

Author information

Author notes

  1. Jean-Michel Masson: Corresponding author.

Authors and Affiliations

  1. INSA, Centre de Transfert en Biotechnologie-Microbiologie, UA 544 du CNRS, Avenue de Rangueil, 31077, Toulouse Cedex, France

    Eliane Meilhoc & Jean-Michel Masson

  2. Centre de Recherche de Biochimie et Génétique Cellulaires du CNRS (UPR 27), Route de Narbonne, 31062, Toulouse, Cedex, France

    Justin Teissié

Authors
  1. Eliane Meilhoc
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jean-Michel Masson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Justin Teissié
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Meilhoc, E., Masson, JM. & Teissié, J. High Efficiency Transformation of Intact Yeast Cells by Electric Field Pulses. Nat Biotechnol 8, 223–227 (1990). https://doi.org/10.1038/nbt0390-223

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0390-223

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing