利用原核表达载体pET-32a构建盐藻磷酸果糖激酶DsPFK基因的表达载体pET32a-DsPFK.转化大肠杆菌BL21(DE3),通过不同温度及不同浓度IPTG诱导后获得以包涵体形式表达的DsPFK重组蛋白,经亲和层析获得大小44.4 kDa的纯化蛋白,通过柱上复性的方法测定重组DsPFK蛋白的酶活为950U.
In this study , vector pET-32 is used to construct the D. salina PFK gene's prokaryotic ex- pression vector pET-32a-DsPFK. Then DsPFK is expressed in Escherichia coli BL21 (DE3), 44kDa recombinant protein is expressed through IPTG induction in inclusion body, the enzyme activity of DsPFK is 950U and determined by post renaturation.