Del Rosario M, Stephans J C, Zakel J, Escobedo J, Giese K
Chiron Technologies, Chiron Corporation, Emeryville, CA 94608, USA.
Nat Biotechnol. 1996 Nov;14(11):1592-6. doi: 10.1038/nbt1196-1592.
We describe a transcription-based assay system to screen for antiviral drugs in vivo. The system consists of two transcription units, a cytomegalovirus promoter driving a reporter gene physically linked to an HIV-1 promoter oriented in the opposite direction. Based on the arrangement of the transcription units, enhanced HIV-1 promoter activity in the presence of the viral transactivating Tat protein downregulates reporter gene expression initiated from the CMV promoter. Inhibitors of HIV-1 transcription relieve the suppression and are identified by an increase in reporter gene expression. This positive selection system allows discrimination between drugs that nonspecifically block cellular functions by cytotoxicity and molecules that specifically repress HIV-1 promoter activity.
我们描述了一种基于转录的体内抗病毒药物筛选检测系统。该系统由两个转录单元组成，一个巨细胞病毒启动子驱动一个报告基因，该报告基因与一个方向相反的HIV-1启动子物理相连。基于转录单元的排列，在病毒反式激活Tat蛋白存在的情况下，增强的HIV-1启动子活性会下调由CMV启动子起始的报告基因表达。HIV-1转录抑制剂可缓解这种抑制作用，并通过报告基因表达的增加来鉴定。这种阳性选择系统能够区分通过细胞毒性非特异性阻断细胞功能的药物和特异性抑制HIV-1启动子活性的分子。
