Shoji I, Aizaki H, Tani H, Ishii K, Chiba T, Saito I, Miyamura T, Matsuura Y
Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
J Gen Virol. 1997 Oct;78 ( Pt 10):2657-64. doi: 10.1099/0022-1317-78-10-2657.
A baculovirus (Autographa californica nucleopolyhedrovirus) vector containing a strong promoter, the CAG promoter, was developed to introduce foreign genes into mammalian cells. Recombinant baculoviruses carrying a reporter gene under the control of the CAG promoter were inoculated into various mammalian cell lines. High-level expression was observed not only in hepatocytes but also in other non-hepatic cell lines tested. Expression of the reporter gene was detected even 14 days after infection. The infectious titre of the recovered baculoviruses decreased significantly after infection, indicating that the baculoviruses did not replicate in mammalian cells. We then compared the efficiencies of gene expression by the baculovirus vector with that of a replication-defective adenovirus vector by using the same expression unit. The same level of expression was observed in HepG2, HeLa and COS7 cells by both vectors. Efficient expression and proper processing were observed in mammalian cells infected with baculoviruses carrying genes coding for structural regions of hepatitis C virus. These results suggest that the baculovirus vector is a good tool for gene delivery into various mammalian cells in order to study the function of foreign genes.
一种含有强启动子（CAG启动子）的杆状病毒（苜蓿银纹夜蛾核型多角体病毒）载体被开发用于将外源基因导入哺乳动物细胞。携带在CAG启动子控制下的报告基因的重组杆状病毒被接种到各种哺乳动物细胞系中。不仅在肝细胞中观察到了高水平表达，在测试的其他非肝细胞系中也观察到了高水平表达。甚至在感染后14天仍检测到报告基因的表达。回收的杆状病毒的感染滴度在感染后显著下降，这表明杆状病毒在哺乳动物细胞中不复制。然后，我们通过使用相同的表达单元，比较了杆状病毒载体与复制缺陷型腺病毒载体的基因表达效率。在HepG2、HeLa和COS7细胞中，两种载体观察到相同水平的表达。在用携带编码丙型肝炎病毒结构区基因的杆状病毒感染的哺乳动物细胞中观察到了高效表达和正确加工。这些结果表明，杆状病毒载体是将基因导入各种哺乳动物细胞以研究外源基因功能的良好工具。
