Targeted integrative transformation ofCandida tropicalisby electroporation

✍ Scribed by Tracy L. Rohrer; Stephen K. Picataggio

• Publisher: Springer
• Year: 1992
• Tongue: English
• Weight: 762 KB
• Volume: 36
• Category: Article
• ISSN: 1432-0614
• DOI: 10.1007/bf00183243

✦ Synopsis

A method for integrative transformation of the diploid yeast Candida tropicalis by electroporation has been developed. By linearizing the transforming plasmid DNA containing the URA3 gene prior to electroporation of recipient cells, its integration was targeted to a specific locus in the genome, resulting in single or multiple tandem integrations. The optimal electroporation conditions for this yeast were established • and include an electric pulse of 2.25 kV/cm for a duration of 50 ms. Using these conditions, Ura + transformants were readily obtained at a high frequency (45 transformants/txg DNA) as the result of targeted integration of the URA3 gene containing plasmid DNA at the chromosomal ura3 locus. The number of transformants resulting from this procedure is comparable to that achieved with a recently reported spheroplast transformation procedure for C. tropicalis; in addition, it offers the advantages of being simple, rapid and reproducible.