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.
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References
Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci U S A 75:1929–1933
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Costanzo MC, Fox TD (1988) Transformation of yeast by agitation with glass beads. Genetics 120:667–670
Simon JR, McEntee K (1989) A rapid and efficient procedure for transformation of intact Saccharomyces cerevisiae by electroporation. Biochem Biophys Res Commun 164:1157–1164
Becker DM, Guarente L (1991) High-efficiency transformation of yeast by electroporation. Methods Enzymol 194:182–187
Delorme E (1989) Transformation of Saccharomyces cerevisiae by electroporation. Appl Environ Microbiol 55:2242–2246
Meilhoc E, Masson JM, Teissie J (1990) High efficiency transformation of intact yeast cells by electric field pulses. Biotechnology (N Y) 8:223–227
Grey M, Brendel M (1992) A ten-minute protocol for transforming Saccharomyces cerevisiae by electroporation. Curr Genet 22:335–336
Rech EL, Dobson MJ, Davey MR, Mulligan BJ (1990) Introduction of a yeast artificial chromosome vector into Saccharomyces cerevisiae cells by electroporation. Nucleic Acids Res 18:1313
Gietz RD, Woods RA (2001) Genetic transformation of yeast. BioTechniques 30:816–820. 22-6, 28 passim
Suga M, Hatakeyama T (2003) High-efficiency electroporation by freezing intact yeast cells with addition of calcium. Curr Genet 43:206–211
Thompson JR, Register E, Curotto J, Kurtz M, Kelly R (1998) An improved protocol for the preparation of yeast cells for transformation by electroporation. Yeast 14:565–571
Jacob HE, Forster W, Berg H (1981) Microbiological implications of electric field effects. II. Inactivation of yeast cells and repair of their cell envelope. Z Allg Mikrobiol 21:225–233
Ganeva V, Galutzov B, Teissie J (1995) Electric field mediated loading of macromolecules in intact yeast cells is critically controlled at the wall level. Biochim Biophys Acta 1240:229–236
Ganeva V, Galutzov B, Teissie J (2014) Evidence that pulsed electric field treatment enhances the cell wall porosity of yeast cells. Appl Biochem Biotechnol 172:1540–1552
Martinez JM, Delso C, Aguilar D, Cebrian G, Alvarez I, Raso J (2018) Factors influencing autolysis of Saccharomyces cerevisiae cells induced by pulsed electric fields. Food Microbiol 73:67–72
Weaver JC, Harrison GI, Bliss JG, Mourant JR, Powell KT (1988) Electroporation: high frequency of occurrence of a transient high-permeability state in erythrocytes and intact yeast. FEBS Lett 229:30–34
Gift EA, Weaver JC (1995) Observation of extremely heterogeneous electroporative molecular uptake by Saccharomyces cerevisiae which changes with electric field pulse amplitude. Biochim Biophys Acta 1234:52–62
Ganeva V, Galutzov B, Teissie J (1995) Fast kinetic studies of plasmid DNA transfer in intact yeast cells mediated by electropulsation. Biochem Biophys Res Commun 214:825–832
Neumann E, Kakorin S, Tsoneva I, Nikolova B, Tomov T (1996) Calcium-mediated DNA adsorption to yeast cells and kinetics of cell transformation by electroporation. Biophys J 71:868–877
Cemazar M, Sersa G, Frey W, Miklavcic D, Teissie J (2018) Recommendations and requirements for reporting on applications of electric pulse delivery for electroporation of biological samples. Bioelectrochemistry 122:69–76
Costaglioli P, Meilhoc E, Masson JM (1994) High-efficiency electrotransformation of the yeast Schwanniomyces occidentalis. Curr Genet 27:26–30
Talmont F, Sidobre S, Demange P, Milon A, Emorine LJ (1996) Expression and pharmacological characterization of the human mu-opioid receptor in the methylotrophic yeast Pichia pastoris. FEBS Lett 394:268–272
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This research was conducted with the support of the EBAM European Associated Laboratory (LEA).
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Meilhoc, E., Teissie, J. (2020). Electrotransformation of Saccharomyces cerevisiae. In: Li, S., Chang, L., Teissie, J. (eds) Electroporation Protocols. Methods in Molecular Biology, vol 2050. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9740-4_21
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DOI: https://doi.org/10.1007/978-1-4939-9740-4_21
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