CLONING OF THE CANDIDA-GLABRATA TRP1 AND HIS3 GENES, AND CONSTRUCTIONOF THEIR DISRUPTANT STRAINS BY SEQUENTIAL INTEGRATIVE TRANSFORMATION

The Candida glabrata (Cg) TRP1 and HIS3 genes have been isolated by co mplementation of the Saccharomyces cerevisiae (Sc) trp1 and his3 mutan ts, respectively. Cg TRP1 encodes a polypeptide of 217 amino acids (aa ), whose aa sequence is 58% identical to that of Sc TRP1. Cg HIS3 enco des a polypeptide of 210 aa, whose aa sequence is 73% identical to tha t of the Sc HIS3. Both Cg TRP1 and HIS3 were disrupted by sequential i ntegrative transformation where the Sc URA3 was used as a selection ma rker for transformation. The resulting auxotrophic strain of his3(-) a nd trp1(-) was used to examine the ability of the Sc genes to compleme nt the Cg mutations; Sc HIS3 and TRP1 complemented the Cg his3(-) and trp1(-) mutations, respectively.