中国水仙类黄酮-3-O-葡糖基转移酶基因的原核表达

Prokaryotic Expression of 3 GT Gene from Narcissus tazetta var. Chinensis

摘要: 通过双酶切、连接转化等方法将中国水仙类黄酮-3-O-葡糖基转移酶（NT3GT）基因克隆到原核表达载体pET29a上，构建原核表达重组质粒pET29a-NT3GT。重组质粒转化至E.coli BL21（DE3）后经IPTG诱导，SDS-PAGE分析表明，经IPTG诱导，NT3GT基因在大肠杆菌BL21（DE3）中获得了高效表达，表达的融合蛋白分子量约为55 kDa。成功构建其原核表达载体并使其在大肠杆菌中得到高效表达，为NT3GT抗血清的制备及功能分析奠定基础。

Abstract: Flavonoid 3-O-glucosyltransferase(Nt3GT) gene of Narcissus tazetta var. Chinensis was cloned into the vector, pET29a, to construct prokaryotic expression recombinant plasmid, pET29a-NT3GT.The recombinant plasmid was transferred to E.coli BL21(DE3) to be induced by IPTG. Subsequently, the expression of the target protein was verified by SDS-PAGE. Then, pET29a-Nt3GT was successfully constructed, which induced the 55 kDa recombinant protein of Nt3GT in the prokaryotic expression system. Nt3GT was cloned, and its vector was constructed and induced to be expressed successfully by IPTG in BL21 providing a basis for theantiserum preparation and functional analysis in the future.