人参皂苷-α-鼠李糖苷酶基因的亚克隆及表达

Sub-cloning and expression of ginsenoside-α-rhanmosidase gene

将人参皂苷-α-鼠李糖苷酶基因亚克隆到毕赤酵母表达载体pPIC9K,再将构建后的表达载体电转化到毕赤酵母GS115中,并诱导表达该基因。结果显示,重组酵母菌在甲醇体积分数为0.5%,72 h培养后,所提取的酶蛋白经酶活性检测,发现具有人参皂苷-α-鼠李糖苷酶的活性,经SDS-PAGE分析,确定其分子质量为52 ku,与理论值基本相符,表明该基因得到了表达。

Gene ginsenoside-α-rhamnosidase was subcloned into an expression vector pPIC9K, the Pichia pastoris GSl15 cells were transformed with the recombinant plasmid by electroporation and expressed. After induction for 72 hours and adding 0.5% methanol, it was showed that the target protein was expressed in Pichia pastoris GSl15 by analyses of the TLC and SDS-PAGE.