van Poelwijk F, Broer R, Belsham G J, Oudshoorn P, Vlak J M, Goldbach R W
Department of Virology, Binnenhaven, The Netherlands.
Biotechnology (N Y). 1995 Mar;13(3):261-4. doi: 10.1038/nbt0395-261.
A hybrid recombinant baculovirus-bacteriophage T7 expression system was developed for transient expression in insect cells of plasmids with foreign genes provided with a T7 promoter. The coding sequence for T7 RNA polymerase, with or without a nuclear localization signal, was inserted into the genome of Autographa californica nuclear polyhedrosis virus. Recombinant viruses stably expressed T7 RNA polymerase in insect cells. Upon transfection of infected insect cells with plasmids containing the genes for chloramphenicol acetyltransferase (CAT), the hepatitis B virus precore-, core- or e- antigens under control of the T7 promoter, transient expression of these genes was detected by ELISA. The results obtained indicate that this baculovirus/T7 system provides a simple and widely applicable tool for transient gene expression studies.
开发了一种混合重组杆状病毒-噬菌体T7表达系统，用于在昆虫细胞中瞬时表达带有T7启动子的含外源基因的质粒。将带有或不带有核定位信号的T7 RNA聚合酶编码序列插入苜蓿银纹夜蛾核型多角体病毒基因组中。重组病毒在昆虫细胞中稳定表达T7 RNA聚合酶。用含有氯霉素乙酰转移酶（CAT）基因、在T7启动子控制下的乙肝病毒前核心抗原、核心抗原或e抗原的质粒转染受感染的昆虫细胞后，通过ELISA检测到这些基因的瞬时表达。所得结果表明，该杆状病毒/T7系统为瞬时基因表达研究提供了一种简单且广泛适用的工具。
