Transformation of Candida albicans by electroporation

• Published in: Yeast (Chichester, England) England), 1999-11, Vol.15 (15), p.1609-1618
• Main Authors: De Backer, Marianne D., Maes, Dirk, Vandoninck, Sandy, Logghe, Marc, Contreras, Roland, Luyten, Walter H. M. L.
• Format: Article
• Language: English
• Subjects: Blotting, Southern; Candida albicans; Candida albicans - chemistry; Candida albicans - genetics; DNA Primers; DNA, Fungal - chemistry; electroporation; Electroporation - methods; episomal vector; homologous recombination; Image Processing, Computer-Assisted; integrative vector; Lithium Compounds - chemistry; Plasmids - chemistry; Polymerase Chain Reaction; Spheroplasts - chemistry; Spheroplasts - genetics; transformation; Transformation, Genetic
• ISSN: 0749-503X; 1097-0061
• DOI: 10.1002/(SICI)1097-0061(199911)15:15<1609::AID-YEA485>3.0.CO;2-Y

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/µg DNA), while spheroplast transformation, although more efficient (∼300 transformants/µg integrative DNA and 103–104 transformants/µg 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/µg were obtained for an integrative plasmid and up to 4500 transformants/µg 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. Copyright © 1999 John Wiley & Sons, Ltd.