重组人卵泡刺激素在昆虫细胞中的表达

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pVLl393，我们得到了表达载体pVLl393-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-hCGoL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。

Expression of human follicle stimulating hormone in insect cells]

To study the expression of hFSH in insect cells, the cDNA encoding the hFSH beta chain was cloned by overlapping-PCR using human chromosome DNA extracted from placental tissue as template. Then we constructed expression vector pVL1393/hFSH beta 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 beta were collected. The beta subunit of hFSH expressed in plasma of the SF9 cells was detected by Western blot analysis, and showed apparent molecular masses of 21 kDa. After coinfecting SF9 cells with recombinant viruses AcNPV-hFSH beta and AcNPV-hCG alpha, secreted heterodimer of hFSH was detected by Western blot under non-reducing conditions. The apparent molecular weight of heterodimer was about 33 kDa.