A PCR-based approach to sequence the Candida tropicalis HSP90 gene.
The heat shock protein 90 (hsp90) gene sequence is known to be highly conserved across the species barrier. A PCR-based method was thus utilised in an attempt to sequence the Candida tropicalis hsp90 gene. Primers for PCR were designed from conserved regions of the gene, which were identified by comparing the Saccharomyces cerevisiae and Candida albicans hsp90 gene sequences. Different sets of primers were designed to amplify and obtain overlapping DNA sequences of the C tropicalis gene. PCR was carried out on genomic DNA of Candidca tropicalis and the PCR products were cloned into suitable vector molecules for sequencing. In this way, a 2,070-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 compared to more conventional methods.
Santhanam
J
J
Department of Medical Microbiology, Manchester Royal Infirmary, UK. jacinta@medic.ukm.my
Burnie
J P
JP
D000595:Amino Acid Sequence; D002175:Candida / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D005800:Genes, Fungal; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004271:DNA, Fungal; D018841:HSP90 Heat-Shock Proteins