Analysis of retroviral assembly using a vaccinia/T7-polymerase complementation system.
Analysis of retroviral assembly using a vaccinia/T7-polymerase complementation system.
复制标题
使用牛痘/T7聚合酶互补系统分析逆转录病毒组装。
DOI：
10.1006/viro.1993.1249
复制
发表时间：
1993
期刊：
影响因子：
3.7
通讯作者：
Hunter,E
中科院分区：
文献类型：
--
作者：
Dong,J;Hunter,E
逆转录病毒组装过程中蛋白质-蛋白质相互作用的性质尚不清楚，并且对病毒组装过程中涉及的潜在信号进行突变分析很困难，特别是对于禽类逆转录病毒，因为在含有缺陷病毒基因组的克隆细胞系中表达的病毒蛋白质水平较高。我们在此描述了一种互补系统，其中逆转录病毒gag/polandenvgene产物在禽类细胞中在bacteriopbage T7启动子的控制下独立于不同质粒表达。来自牛痘病毒载体的 T7 聚合酶的共表达导致逆转录病毒结构蛋白的高水平生物合成和病毒颗粒的有效组装。电子显微镜和蛋白质成分分析表明，这些病毒粒子与 RSV 感染细胞产生的病毒粒子没有区别。通过使用突变糖蛋白基因，可以证明组装过程的特异性，并描述了该系统对其他逆转录病毒系统的适用性。
The nature of protein-protein interactions during retroviral assembly is not well understood, and mutational analyses of the potential signals involved in the viral assembly process has been difficult, particularly with the avian retroviruses due to the level of viral proteins expressed in the clonal cell lines containing defective viral genomes. We describe here a complementation system in which the retroviralgag/polandenvgene products were expressed independently from different plasmids under the control of the bacteriopbage T7 promoter, in avian cells. Coexpression of the T7 polymerase from a vaccinia virus vector resulted in a high level of biosynthesis of retroviral structural proteins and efficient assembly of virus particles. Electron microscopy and protein composition analyses demonstrated that these virions were indistinguishable from those produced from RSV-infected cells. Through the use of mutant glycoprotein genes it was possible to demonstrate the specificity of the assembly process and the applicability of this system to other retroviral systems is described.