Fujita Mikako, Matsumoto Sora, Sakurai Akiko, Doi Naoya, Miyaura Maki, Yoshida Akiko, Adachi Akio
Department of Virology, The University of Tokushima Graduate School of Medicine, Tokushima-shi, Tokushima 770-8503, Japan.
Microbes Infect. 2002 Oct;4(12):1203-7. doi: 10.1016/s1286-4579(02)01646-5.
The vif gene of human immunodeficiency virus type 1 (HIV-1) is essential for virus growth in non-permissive cells such as H9. To elucidate the mechanism of action of the Vif protein, vif mutants, which show trans-dominant negative effects on the replication of HIV-1, would be useful tools. In this study, a new assay system to identify the mutants of this category was established. For this new system, various reporter clones carrying both mutant and authentic vif sequences were generated. By determining the growth ability of the viruses derived from the reporter constructs, the potential negative effect of the mutant vif sequence was readily and sensitively monitored. Ten vif mutant sequences tested were found not to exert the trans-dominant negative effect on the replication of HIV-1.
人类免疫缺陷病毒1型(HIV-1)的vif基因对于病毒在非允许细胞(如H9细胞)中的生长至关重要。为阐明Vif蛋白的作用机制,对HIV-1复制具有反式显性负效应的vif突变体将是有用的工具。在本研究中,建立了一种新的检测系统来鉴定此类突变体。对于这个新系统,构建了携带突变型和野生型vif序列的各种报告克隆。通过测定源自报告构建体的病毒的生长能力,可以轻松且灵敏地监测突变型vif序列的潜在负效应。所测试的10个vif突变序列未发现对HIV-1复制产生反式显性负效应。