论文部分内容阅读
目的:构建含有ULBP1基因启动子的报告质粒,并初步研究NS3/4A对ULBP1转录的影响。方法:将ULBP1启动子核心区序列连接在pGL3-enhance载体上构建ULBP1报告质粒pGL3-ULBP1质粒;将HCV的蛋白酶NS3/4A基因亚克隆到pcDNA3-Flag载体上(pcDNA3-Flag-NS3/4A),Western blot检测其表达。用荧光分度计检测NS3/4A对ULBP1转录水平的影响。结果:成功构建了含有ULBP1启动子的报告质粒pGL3-ULBP1和FLAG-NS3/4A真核表达质粒,并检测到NS3/4A可以抑制ULBP1的转录。结论:HCV的蛋白酶NS3/4A可以抑制ULBP1的转录。
OBJECTIVE: To construct a reporter plasmid containing ULBP1 promoter and to study the effect of NS3 / 4A on ULBP1 transcription. METHODS: ULBP1 promoter core region sequence was ligated to pGL3-enhance vector to construct ULBP1 reporter plasmid pGL3-ULBP1 plasmid; HCV protease NS3 / 4A gene was subcloned into pcDNA3-Flag vector (pcDNA3-Flag-NS3 / 4A) Western blot was used to detect the expression. The effect of NS3 / 4A on the ULBP1 transcription level was detected by fluorescence spectrophotometer. Results: The eukaryotic expression plasmids of pGL3-ULBP1 and FLAG-NS3 / 4A containing ULBP1 promoter were successfully constructed and NS3 / 4A could inhibit the transcription of ULBP1. Conclusion: HCV protease NS3 / 4A can inhibit the transcription of ULBP1.