Abstract:

The aim was to obtain a constitutive promoter for construction of expression vector of industrial Saccharomyces cerevisiae. Applying S. cerevisiae NYK genomic DNA as template, two fragments upstream the initiate codes gene were obtained by PCR. The long fragment was 781 bp and designated PGK1 (GenBank Accession No.FJ415226). Sequence analysis by NCBI Blast showed that the nucleotide sequence of PGK1 was 99% homologous to PGK promoter located in chromosome Ⅲ from S. cerevisiae (GenBank Accession No X59720). TATA box, CAAT-box and other regulatory elements for gene expression were detected in the nucleotide sequence of PGK1. Functional analysis indicated that PGK1 could initiate the expression of the foreign glucoamylase gene (ga1) integrated in industrial S. cerevisiae. The results showed that promoter PGK1 was obtained which provided experimental base for construction of expression vector of industrial S. cerevisiae.