Electrotransformation of Saccharomyces cerevisiae

Methods Mol Biol. 2020:2050:187-193. doi: 10.1007/978-1-4939-9740-4_21.

Abstract

Intact yeast cell transformation is easily achieved by gene electrotransfer (GET). The procedure is fast and efficient in terms of transformants/μg DNA. Yeast cells in exponential growth phase are washed, treated for a short period with dithiothreitol (DTT) and then mixed with the plasmid DNA in a buffer with a low conductivity. A single well defined electric pulsed is delivered. After a 1 h incubation in the growth medium without selection, transformants are obtained on a selective plate medium. After a short description of the present knowledge on the events affecting the yeast cell as a consequence of the pulsed electric field, a step-by-step protocol is reported for Saccharomyces cerevisiae.

Keywords: Electrotransformation; Gene electrotransfer; Saccharomyces; Yeast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dithiothreitol / pharmacology
  • Electroporation / methods*
  • Microbiological Techniques
  • Plasmids / genetics*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development*
  • Transformation, Genetic

Substances

  • Dithiothreitol