Hou Li Li, Jiang Shu, Shen Wei Ying, Huang Zhe Ping, Shen Qing Xiang
Shanghai Institute of Planned Parenthood Research, Shanghai 200032.
Shi Yan Sheng Wu Xue Bao. 2004 Apr;37(2):133-8.
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.