Ochsenbauer C, Wilk T, Bosch V
Forschungsschwerpunkt Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
J Gen Virol. 1997 Mar;78 ( Pt 3):627-35. doi: 10.1099/0022-1317-78-3-627.
In order to facilitate analyses of the molecular function of the human immunodeficiency virus type 1 (HIV-1) Vif protein, we have developed a cell culture model-system which allows permanent production of genotypically and phenotypically vif-defective HIV-1 virions in 'non-permissive' H9 cells. Using recombinant, replication-competent HIV-1 proviruses coding for a selectable marker gene (gpt) instead of nef, two stably infected H9 subclones named M2 (vif-mutant) and WX (wild-type), respectively, were generated. Virions released from cell line M2--displaying the expected vif-defective phenotype--are produced permanently, and in an at least 50 times higher amount than virus particles from acutely vif-negative HIV-1-infected H9 cells. Analysis of viral protein composition and the electron-microscopic morphology of vif-mutant virions did not reveal any detectable differences in comparison to wild-type virions.
为便于分析人类免疫缺陷病毒1型(HIV-1)Vif蛋白的分子功能,我们建立了一种细胞培养模型系统,该系统可在“非允许性”H9细胞中持续产生基因和表型上Vif缺陷的HIV-1病毒粒子。利用编码可选择标记基因(gpt)而非nef的重组、具有复制能力的HIV-1前病毒,分别产生了两个稳定感染的H9亚克隆,命名为M2(Vif突变体)和WX(野生型)。从细胞系M2释放的病毒粒子呈现出预期的Vif缺陷表型,可持续产生,且产生量至少比急性Vif阴性HIV-1感染的H9细胞产生的病毒颗粒高50倍。与野生型病毒粒子相比,对Vif突变体病毒粒子的病毒蛋白组成和电子显微镜形态分析未发现任何可检测到的差异。