High-efficiency electrotransformation of the yeast Schwanniomyces occidentalis.
A method has been developed for introducing heterologous DNA rapidly and efficiently by electropermeabilization into the yeast Schwanniomyces occidentalis. A transformation efficiency as high as 2 x 10(5) transformants/microgram of plasmid DNA was obtained with a square-wave electric pulse of 2.17 kV/cm during 18 ms. Small quantities of DNA (5 ng) can be used to transform 3 x 10(8) cells. The main parameters which have been optimized are: presence of adenine in the culture medium, pretreatment of the cells with dithiothreitol during the exponential growth phase of the cells, amount of cells treated, and pulse-field strength and duration. Competent cells can be stored to allow electrotransformation whenever needed.
Costaglioli
P
P
Institut National des Sciences Appliquées, CNRS, Toulouse, France.
Meilhoc
E
E
Masson
J M
JM
D000225:Adenine / Q000494:pharmacology; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D004271:DNA, Fungal / Q000235:genetics; D004229:Dithiothreitol / Q000494:pharmacology; D018274:Electroporation; D009994:Osmolar Concentration; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic / Q000187:drug effects; D015003:Yeasts / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development
D003451:Cryoprotective Agents; D003470:Culture Media; D004271:DNA, Fungal; D000225:Adenine; D004229:Dithiothreitol