A PCR-based approach to sequence the Candida tropicalis HSP90 gene

The heat shock protein 90 (hsp90) gene sequence is known to be highly conse rved across the species barrier. A PCR-based method was thus utilised in an attempt to sequence the Candida tropicalis hsp90 gene. Primers for PCR wer e designed from conserved regions of the gene, which were identified by com paring the Saccharomyces cerevisiae and Candida albicans hsp90 gene sequenc es. Different sets of primers were designed to amplify and obtain overlappi ng DNA sequences of the C. tropicalis gene. PCR was carried out on genomic DNA of Candida tropicalis and the PCR products were cloned into suitable ve ctor molecules For sequencing. In this way, a 2070-basepair sequence of the C. tropicalis hsp90 gene was obtained. The PCR-based approach proved to be an easier method of obtain the sequence of a highly conserved gene, as com pared to more conventional methods. (C) 2000 Federation of European Microbi ological Societies. Published by Elsevier Science B.V. All rights reserved.