酵母α-半乳糖苷酶组成型表达基因的重组构建及转化

Construction and Transformation of A Constitutive Expression α-Galactosidase Gene in Yeast

用 PCR 方法从酿酒酵母 AH109 中扩增出 α-半乳糖苷酶 MEL1 基因片段,与乙醇脱氢酶组成型启动子重组后构建成表达重组质粒 pGAD-GAL.将重组质粒 pGAD-GAL 转化不带 α-半乳糖苷酶基因的酿酒酵母后,在预先涂布有 X-α-Gal 的亮氨酸缺陷培养基(SD/-Leu)平板上选择到蓝色菌落.实现了 α-半乳糖苷酶在酵母内的组成型表达.

MEL1 gene was amplified in Saccharomyces cerevisiae AH109 with PCR,and Recombined with ADH1 constitutive promotor thus recombinant plasmid pGAD-GAL was constructed.The recombinant plasmid pGAD-GAL was transformed into yeast,and the blue yeast clonies was screened out on SD/-Leu nutritional media with X-alpha-Gal.Blue colonies indicated the alpha -galactosidase was constitutively expression in Saccharomyces cerevisiae.