Gibellini D, Re M C, Panaya R, Venturi E, Milani D, La Placa M, Zauli G
Department of Clinical and Experimental Medicine, University of Bologna, Italy.
J Immunol Methods. 1998 Dec 1;221(1-2):107-17. doi: 10.1016/s0022-1759(98)00169-0.
The transactivator Tat protein represents a pivotal factor for the replication of human immunodeficiency virus type 1 (HIV-1). In this report, we describe a flow cytometry procedure designed to quantify the intracellular content of Tat protein in Jurkat CD4+ T lymphoblastoid cell lines, stably transfected with plasmids expressing full-length Tat protein. Various expression vectors were compared for their effectiveness to yield Tat protein in Jurkat cells, and several technical parameters were analyzed to optimize the assay. This method offers a quick and efficient approach to select stably transfected cell lines expressing different levels of specific protein.
反式激活蛋白Tat是1型人类免疫缺陷病毒（HIV-1）复制的关键因子。在本报告中，我们描述了一种流式细胞术方法，用于定量分析稳定转染了表达全长Tat蛋白质粒的Jurkat CD4+ T淋巴母细胞系中Tat蛋白的细胞内含量。比较了各种表达载体在Jurkat细胞中产生Tat蛋白的有效性，并分析了几个技术参数以优化该检测方法。该方法为筛选表达不同水平特定蛋白的稳定转染细胞系提供了一种快速有效的途径。
