【摘要】 为分析JDV与BIV、HIV-1LTR和Tat相互激活能力差异的原因,在氨基酸序列对比及HIV-1Tat功能域划分的基础上构建了JH、HJ、JB、BJ几种嵌合Tat蛋白,并克隆到真核表达载体。将上述表达质粒与以JDV、BIV和HIV-1LTR为启动子,以luc为报告基因的质粒共转染Hela细胞,证实了三种不同Tat激活能力的差异主要来自其结合域RNA结合能力的差异,排除了结构域不完整和细胞因子缺乏造成JH不激活HIV-1LTR的可能性。更多还原

【Abstract】 In order to find the cause of different transactivate abilities among JDV, BIV and HIV-1 Tat, four chimeric Tat cDNA expression constructs, JH, HJ, JB and BJ, were generated with the crossover points at the boundary of activation and binding domains based on functional domain division of HIV-1 Tat and amino acid sequence comparision among HIV-1, JDV and BIV Tat. These chimeric Tat proteins were co-transfected with JDV, BIV and HIV-1 LTR report plasmids in Hela. Transient assay showed that different transactivate abilities among JDV, BIV and HIV-1 Tat were mainly caused by different binding abilities of their binding domains. We further excluded some possibilities that may cause the poor transactivate ability of JH on HIV-1 LTR, such as incomplete functional domains and the lack of related cytokines.更多还原