High-efficiency transformation of the pathogenic yeast Candida parapsilosis.
A recently developed transformation system for the pathogenic yeast Candida parapsilosis opened a venue for studying the biological phenomena of this species at the molecular level. However, the standard chemical method yielded only about 1x10(3) transformants/microg of DNA, which is insufficient for certain types of experiment. With the aim of increasing the transformation efficiency, we employed two alternative methods for the introduction of plasmids into the recipient cells. Whereas biolistics resulted in about 5x10(2) transformants/microg of plasmid DNA, electroporation was an order of magnitude more efficient than the chemical method. Pretreatment of cells with 100 mM lithium acetate or 10 mM dithiothreitol resulted in a 5-fold (5x10(4)) or a 10-fold (1x10(5)) increase in transformation efficiency, respectively. This high-efficiency transformation method should be suitable for experiments such as the screening of DNA libraries.
Zemanova
Julia
J
Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Mlynská dolina CH-1, 84215, Bratislava, Slovakia. julia.zemanova@fns.uniba.sk
Nosek
Jozef
J
Tomaska
Lubomir
L
D000085:Acetates / Q000494:pharmacology; D019470:Biolistics; D002175:Candida / Q000235:genetics* / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D018274:Electroporation; D015723:Gene Library; D005820:Genetic Testing; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics* / Q000502:physiology
D000085:Acetates; C488804:lithium acetate; D004229:Dithiothreitol