Karschin A
Division of Biology, California Institute of Technology, Pasadena 91125.
EXS. 1993;63:31-47. doi: 10.1007/978-3-0348-7265-2_2.
Versatile and potent expression systems are needed to decipher the structure and functions of the many excitability proteins that have been identified through molecular cloning. This article reviews the use of recombinant vaccinia viruses (VV), which have been recently explored for the heterologous expression of eukaryotic proteins. Vaccinia viruses feature a series of favourable properties, most of all a broad host range and high efficiency of infection, that make them uniquely suited as flexible expression vectors. In one type of experiment, the recombinant virus simply harbors the cDNA for the foreign protein; in a second type the virus harbors the cDNA for the specific and efficient RNA polymerase of bacteriophage T7, which in turn generates RNA from a separate introduced plasmid or virus. Both variations have been successfully applied to the expression and analysis of voltage-dependent ion channels, neurotransmitter receptors and other excitability proteins in many cell lines and postmitotic cells in culture. VV vectors promise to be particularly useful to study membrane proteins that require posttranslational processing, association with cell-specific subunits or coupling to endogenous second messengers pathways.
需要通用且高效的表达系统来解析通过分子克隆鉴定出的众多兴奋性蛋白的结构和功能。本文综述了重组痘苗病毒（VV）的应用，最近人们对其用于真核蛋白的异源表达进行了探索。痘苗病毒具有一系列有利特性，最重要的是宿主范围广和感染效率高，这使其特别适合作为灵活的表达载体。在一类实验中，重组病毒仅携带外源蛋白的cDNA；在另一类实验中，病毒携带噬菌体T7特异性高效RNA聚合酶的cDNA，该聚合酶进而从单独导入的质粒或病毒产生RNA。这两种变体均已成功应用于许多细胞系和培养中的有丝分裂后细胞中电压依赖性离子通道、神经递质受体及其他兴奋性蛋白的表达和分析。VV载体有望在研究需要翻译后加工、与细胞特异性亚基结合或与内源性第二信使途径偶联的膜蛋白方面特别有用。
