Electroporation of Pichia pastoris
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The introduction of foreign DNA into the yeast Pichia pastoris is similar to the processes used with Saccharomyces cerevisiae and routinely performed by the electroporation of plasmid DNA into electro-competent (E-comp) P. pastoris cells (Becker and Guarente, Methods Enzymol 194:182–187, 1991). Transformation of spheroplasts is more difficult (Cregg et al., Mol Cell Biol 5:3376–3385, 1985), and current methods to make chemically competent P. pastoris cells results in significantly fewer transformants (Pichia protocols, 2nd edn, Totowa, 27–42). Following are protocols to prepare E-comp P. pastoris cells, and to electroporate plasmid DNA into these cells to generate transformed cells.
KeywordsPichia pastoris Transformation Electroporation Competent cells Spheroplasts
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