Lieber A, Kiessling U, Strauss M
Akademie der Wissenschaften der DDR, Zentralinstitut für Molekularbiologie, Berlin-Buch, GDR.
Nucleic Acids Res. 1989 Nov 11;17(21):8485-93. doi: 10.1093/nar/17.21.8485.
Here we describe a novel expression system for mammalian cells which is based on transcription of hybrid genes containing T7 phage promoters by a T7 phage RNA polymerase targeted to the nucleus of the host cells. The RNA polymerase gene of T7 phage has been modified by substituting a sequence encoding the nuclear location signal of SV40 large T antigen for the N-terminal part of the polymerase gene. Expression of the modified gene is driven by the mouse metallothionein promoter in transfected mouse Ltk- cells resulting in high concentration of the polymerase in the nucleus. Nuclear T7 RNA polymerase directs efficient transcription of the cat gene under control of a T7 promoter. T7 constructs are expressed at a level at least 6 fold higher than the prototype pRSVcat. The unique properties of this heterologeous expression system are discussed.
在此，我们描述了一种用于哺乳动物细胞的新型表达系统，该系统基于通过靶向宿主细胞核的T7噬菌体RNA聚合酶对含有T7噬菌体启动子的杂交基因进行转录。T7噬菌体的RNA聚合酶基因已被修饰，即将编码SV40大T抗原核定位信号的序列替换聚合酶基因的N端部分。修饰基因的表达由小鼠金属硫蛋白启动子驱动，转染小鼠Ltk-细胞后，可使细胞核中聚合酶的浓度很高。核T7 RNA聚合酶在T7启动子的控制下指导cat基因的高效转录。T7构建体的表达水平比原型pRSVcat至少高6倍。本文讨论了这种异源表达系统的独特性质。
