禽流感病毒NS1蛋白亚细胞定位研究

Subcellular Localization of Avian Influenza Virus NS1 Protein

为深入研究NS1蛋白的生物学功能,了解NS1蛋白在哺乳动物细胞中的分布和亚细胞定位情况。采用RT-PCR扩增出禽流感病毒NS1基因片段,克隆至载体pEGFP-N1,经酶切和测序鉴定后,将构建的重组质粒pEGFP-N1-NS1经脂质体介导转染293T细胞和Hela细胞,在荧光显微镜和激光扫描共聚焦显微镜下观察NS1在细胞中的分布、亚细胞定位情况。成功构建了真核重组表达质粒pEGFP-N1-NS1,并在293T细胞及Hela细胞中得到高效表达。融合蛋白发出的绿色荧光表明NS1蛋白主要分布在细胞核,在胞浆中有少量表达。本研究成功在哺乳动物细胞中表达NS1蛋白,并对其进行亚细胞定位,为流感病毒的相关功能研究奠定了基础。

In order to deeply study on biological function of NS1 protein,learn about distribution and subcellular localization of NS1 protein in mammalian cells.NS1 gene of avian influenza virus was amplified by RT-PCR,and cloned into vector pEGFP-N1.Recombinant plasmid was identified by enzyme digestion and sequencing,the recombinant plasmid pEGFP-N1-NS1 was transfected into 293T cells and Hela cells by liposome,distribution and subcellular localization of NS1 protein was observed by the fluorescence microscope and the confocal laser scanning microscope.Eukaryotic recombinant expression plasmid pEGFP-N1-NS1 was successfully constructed,it had high levels of expression.in 293T cells and Hela cells.NS1 green fluorescent fusion protein is mainly distributed in the nucleus,is slightly distributed in the cytoplasm.The study successfully expressed NS1 protein in mammalian cells and analyzed subcellular localization of NS1protein.It is a solid basis for related function research on influenza virus.