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[目的]为研究pRRL为基础的慢病毒系统除了作为RNA干扰工具外是否可用于介导外源基因过表达。[方法]构建能表达NALP2蛋白N末端PYRIN结构域(PYD)或p53的慢病毒;采用RT-PCR、Western印迹法检测慢病毒介导的PYD、p53表达情况;采用NF-κB荧光素酶报告实验和细胞凋亡实验鉴定PYD和p53的生物学功能。[结果]通过对绿色荧光蛋白(EGFP)的观察和对PYD、p53表达水平的检测,表明成功构建了能够表达PYD或p53的慢病毒。NF-κB荧光素酶报告实验和细胞凋亡实验结果表明慢病毒表达的PYD和p53具有生物学功能。[结论]pRRL为基础的慢病毒系统除了能够作为RNA干扰工具外,还可介导外源基因的表达,并在体外高效感染肿瘤细胞。
[Objective] To investigate whether pRRL-based lentiviral systems can be used to mediate the overexpression of foreign genes in addition to being an RNA interference tool. [Method] To construct the lentivirus which can express PYRIN domain or PYRIN domain of NALP2 protein. The expression of PYD and p53 were detected by RT-PCR and Western blotting. The expression of PYD and p53 was detected by NF-κB luciferase Experiments and apoptosis assays identify the biological functions of PYD and p53. [Result] The observation of green fluorescent protein (EGFP) and the detection of PYD and p53 expression showed that the lentivirus capable of expressing PYD or p53 was successfully constructed. The results of NF-κB luciferase reporter assay and apoptosis showed that lentivirus-expressing PYD and p53 have biological functions. [Conclusion] The pRRL-based lentiviral system can not only serve as an RNA interference tool, but also can mediate the expression of foreign genes and efficiently infect tumor cells in vitro.