An improved transformation protocol for the human fungal pathogen Candida albicans.
Commonly used protocols for the transformation of the dimorphic human fungal pathogen Candida albicans rely on established methods for the yeast Saccharomyces cerevisiae. With respect to transformation efficiency, however, there is a great difference between these two organisms when using the lithium acetate procedure. Here we present a modified version of this protocol for use with C. albicans. Among the different parameters tested, two turned out to be particularly relevant and, when combined, resulted in an up to 10-fold increase in transformation efficiency (400-500 integrative transformants) compared with previous protocols: first, adjusting the heat shock applied to the cells to 44 degrees C for C. albicans instead of 42 degrees C for S. cerevisiae and, second, treating C. albicans cells with lithium acetate in an overnight incubation instead of for 30 min as used for S. cerevisiae. With these modifications, the lithium acetate procedure becomes a very efficient and reliable tool for C. albicans transformation.
Walther
Andrea
A
Junior Research Group: Growth-Control of Fungal Pathogens, Hans-Knöll Institut für Naturstoff-Forschung, Friedrich-Schiller-University, Winzerlaer Strasse 10, 07745, Jena, Germany.
Wendland
Jürgen
J
D000085:Acetates / Q000378:metabolism; D002176:Candida albicans / Q000235:genetics* / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods; D006358:Hot Temperature; D008094:Lithium / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*
D000085:Acetates; D008094:Lithium