Koken S E, van Wamel J, Berkhout B
Department of Virology, University of Amsterdam, The Netherlands.
Gene. 1994 Jul 8;144(2):243-7. doi: 10.1016/0378-1119(94)90384-0.
We developed a sensitive vector system for the analysis of weak promoter activities. This promoter assay is based on the transcriptional activator protein, Tat, of human immunodeficiency virus type 1 (HIV-1). High-level expression of HIV requires activation in trans by Tat of the promoter in the long terminal repeat (LTR). Here we describe the construction of a promoterless pTat vector. Foreign promoter elements can be inserted upstream from the tat gene, and expression of Tat protein is measured in trans on a co-transfected LTR-CAT reporter plasmid. We show that this binary system is more sensitive than standard pCAT reporter assays.
我们开发了一种用于分析弱启动子活性的灵敏载体系统。该启动子检测法基于人类免疫缺陷病毒1型（HIV-1）的转录激活蛋白Tat。HIV的高水平表达需要Tat对长末端重复序列（LTR）中的启动子进行反式激活。在此，我们描述了一种无启动子pTat载体的构建。可将外源启动子元件插入tat基因的上游，并在共转染的LTR-CAT报告质粒上反式检测Tat蛋白的表达。我们证明，该二元系统比标准的pCAT报告检测法更灵敏。
