Effects of GAL₁₀-SUC₂ promoter combinations on SUC₂ gene expression in S. cerevisiae

Abstract

The plasmid series YEP_{5 1 Δ n} bearing GAL₁₀-SUC₂ promoter combinations were constructed by inserting SUC₂ gene with different upstream deletions downstream GAL₁₀ promoter on YEP_{5 1}. After transforming yeast cells S. cerevisiae, the invertases expressed by eacn of the transformants were measured and analysed by means of PAGE. The results showed that: 1) The SUC₂ gene with upstream deletion to at -636bp expressed high level glycosylated form of invertase under glucose derepression, while SUC₂ gene with more extensive deletions to -223 bp or more lost its response to glucose derepression; 2) Each part of GAL₁₀-SUC₂ promoter combination acted almost independently. All of the combinations showed no apparent coordinated promoter function under our experimental conditions; 3) Sequences between -89bp and -41bp of SUC₂ upstream region are responsible for constitutive expression of nonglycosylated invertase. The two tracts of poly (dA-dT) of this region may serve as promoter elements.

Keywords

SUC₂ gene / GAL₁₀ gene / promoter combination / invertase expression

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