重组人卵泡刺激素在昆虫细胞中的表达  

EXPRESSION OF HUMAN FOLLICLE STIMULATING HORMONE IN INSECT CELLS

作　　者：侯理理[1] 蒋俶[1] 沈卫英[1] 黄哲平[1] 申庆祥[1] 

机构地区：[1]上海市计划生育科学研究所

出　　处：《实验生物学报》2004年第2期133-138,共6页Acta Biologiae Experimentalis Sinica

摘　　要：为了研究在昆虫细胞中表达重组人卵泡刺激素,我们以人胎盘组织提取的染色体DNA为模板,利用重叠PCR方法扩增出hFSHβ亚基的cDNA的编码区。将此cDNA克隆入核型多角体病毒(AcNPV)非融合蛋白基因表达载体pVL1393,我们得到了表达载体pVL1393-hFSHβ,然后与BaculoGold^(TM)线性杆状病毒DNA共转染昆虫细胞SF9,经多次扩增后获得高滴度的重组病毒AcNPV-hFSHβ。将此重组病毒感染昆虫细胞,我们得到了在胞浆中表达的hFSHβ亚基,Western blot显示分子量大约为21kDa。以重组病毒AcNPV-hFSHβ与AcNPV-hCGα一同感染昆虫细胞得到了具有分泌性的重组hFSH异二聚体,在非还原的条件下Western blot显示分子量大约为33kDa。To study the expression of hFSH in insect cells, the cDNA encoding the hFSH β chain was cloned by overlapping-PCR using human chromosome DNA extracted from placental tissue as template. Then we constructed expression vector pVL1393/hFSH β using an unfused protein nuclear polyhedrosis virus (AcNPV) expression vector. The insect cells (SF9) were cotransfected with the expression vector and nuclear polyhedrosis linearized virus DNA, and recombinant viruses AcNPV-hFSH β were collected. The β subunit of hFSH expressed in plasma of the SF9 cells was detected by Western blot analysis, and showed apparent molecular masses of 21kDa. After coin-fecting SF9 cells with recombinant viruses AcNPV-hFSH 3 and AcNPv-hCG α , secreted het-erodimer of hFSH was detected by Western blot under non-reducing conditions. The apparent molecular weight of heterodimer was about 33kDa.

关 键 词：人卵泡刺激素 表达 昆虫细胞 重组病毒 hFSH 糖蛋白 

分 类 号：Q57[生物学—生物化学]