Transformation of Candida albicans by electroporation.
In contrast to a variety of other yeasts, Candida albicans has proved difficult to transform with high efficiency. Lithium acetate transformation is fast and simple but provides a very low efficiency of DNA transfer (50-100 transformants/microg DNA), while spheroplast transformation, although more efficient ( approximately 300 transformants/microg integrative DNA and 10(3)-10(4) transformants/microg replicative DNA), is complicated and time-consuming. In this study we applied various yeast transformation techniques to C. albicans and selected an electroporation procedure for further optimization. Transformation efficiencies of up to 300 transformants/microg were obtained for an integrative plasmid and up to 4500 transformants/microg for a CARS-carrying plasmid. This reasonably high transformation efficiency, combined with the ease and speed of electroporation in comparison to alternative techniques, make it the preferred method for transformation of C. albicans.
De Backer
M D
MD
Department of Advanced Biotechnologies, Janssen Research Foundation, Turnhoutseweg 30, B2340 Beerse, Belgium. mdbacker@janbe.jnj.com
Maes
D
D
Vandoninck
S
S
Logghe
M
M
Contreras
R
R
Luyten
W H
WH
D015139:Blotting, Southern; D002176:Candida albicans / Q000737:chemistry / Q000235:genetics*; D017931:DNA Primers; D004271:DNA, Fungal / Q000737:chemistry; D018274:Electroporation / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D018020:Lithium Compounds / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry; D016133:Polymerase Chain Reaction; D013104:Spheroplasts / Q000737:chemistry / Q000235:genetics; D014170:Transformation, Genetic
D017931:DNA Primers; D004271:DNA, Fungal; D018020:Lithium Compounds