果糖缬氨酸氧化酶在sf9细胞中的表达

Expression of Fructose Valine Oxidase in Sf9 Cells

利用杆状病毒表达系统在昆虫细胞sf9中表达果糖缬氨酸氧化酶基因(FVO)。人工合成FVO基因,与载体pFastBac1连接,转化至大肠杆菌DH10Bac感受态细胞,进行同源重组。经筛选得到重组杆状病毒载体Bacmid-pFastBac1-FVO,PCR鉴定得到单一条带;用该重组载体转染sf9细胞,收获毒种并进行表达;对表达产物进行SDS-PAGE、Western blot检测。成功构建了FVO的杆状病毒表达载体,并在sf9细胞中得到了表达,Western blot检测出现1条约40 kD的带。

It was to express the Fructose Valine Oxidase Gene with baculovirns expression system. FVO gene was artificially synthesized, and ligated to the vector pFastBacl to obtain the recombinant vector, which was then transformed into E.coli DH10Bac competent cells to transposase with bacmid. After screening, the recombinant baculovirus Bacmid-pFastBacl-FVO was got, which appeared a single band after PCR identification, sf9 cells was transfected with the recombinant vector, and harvested the virus and expressed the protein. Results showed that the baculovirus expression vector was successfully constructed and expressed in the sf9 cells which appeared a 40 kD band with Western blot.