High efficiency transformation of intact yeast cells by electric field pulses

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 10(7) transformants/micrograms of plasmid DNA were obtained with a square wave...

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Bibliographic Details
Published inBio/technology (New York, N.Y. 1983) Vol. 8; no. 3; pp. 223 - 227
Main Authors Meilhoc, E. (INSA, France), Masson, J.M, Teissie, J
Format Journal Article
LanguageEnglish
Published United States 01.03.1990
Subjects
ADN
Cell Survival
Cloning, Molecular - methods
Dithiothreitol - pharmacology
DNA
ELECTRICIDAD
ELECTRICITE
ELECTRICITY
GENETIC TRANSFORMATION
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - cytology
Saccharomyces cerevisiae - genetics
Transfection - drug effects
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
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Summary: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 10(7) transformants/micrograms 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 10(8) 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.
Bibliography:9028015
F30
ISSN:0733-222X
1087-0156
2331-3684
1546-1696
DOI:10.1038/nbt0390-223