Production of secretory protein in Nonlytic baculovirus expression system

• Main Authors: Yu-Lung Wu, 吳裕隆
• Other Authors: Yu-Kuo Liu
• Format: Others
• Language: zh-TW
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/91521251703023671437

碩士 === 長庚大學 === 生化與生醫工程研究所 === 92 === The baculovirus expression vector system (BEVS) is widely used for recombinant proteins production. The system has several advantages including high protein yield, faster production, ease of use, and safty . In addition, it can perform most of the post-translational modifications observed in mammalian cells. But it is a lytic system and cell lysis can affect both quality and quantity of the product. Dr. Yu-Chan Chao at the Institute of Molecular Biology, Academia Sinica, constructed a nonlytic baculovirus expression system. We employed the nonlytic system to express secreted protein which is low protein yield in the tradition lytic BEVS. In this experiment we used secreted human placental alkaline phosphatase (SEAP) as reporter gene. The results show nonlytic virus can delay cell lysis, but not to increase the SEAP quantity. However, SEAP protein is a stable protein. It will not degrade by the proteinase released following cell lysis. So it could not show the advantage of nonlytic baculovirus expression system. And we found that nonlytic virus vAC4hRpS has very high infection ability even in suspension culture virus infection. It could expression protein early through the high infection rate. We believe this property will benifit good for unstable protein production in the bioreactor.