Identification of a nuclear pheromone-sensitive protein kinase not identical to p34CDC28 in Saccharomyces cerevisiae

Abstract A fragment containing the full length cDNA from Aspergillus oryzae α-amylase has been amplified by PCR using specific synthetic oligonucleotides. The amplified cDNA was designed to favour its expression in yeast by modifying its upstream untranslated region. It was subcloned in the expression vector pYEXα1, placed under the control of the yeast CYC1-GAL10 promoter and used to transform Saccharomyces cerevisiae . Cells were then able to express and secrete active α-amylase to the medium in a regulated fashion. The recombinant enzyme had similar electrophoretic mobility and catalytic properties to the original A. oryzae α-amylase.