禽流感病毒H5N1亚型NS1蛋白的真核表达及其在细胞中的分布

Eukaryotic expression and localization of NS1 protein of H5N1 avian influenza virus in infected cells

为构建禽流感病毒(AIV)H5N1亚型非结构蛋白NS1的真核表达载体,并鉴定其在哺乳动物细胞中的表达与分布,本研究采用RT-PCR技术,从甲型流感病毒的总RNA中扩增NS1全长基因,并将其克隆于pXJ40中,构建真核表达载体pXJ40-HA-NS1。将该重组质粒转染293T细胞,通过western blot方法鉴定表达的NS1蛋白;并以免疫荧光技术观察NS1在H1299细胞中的分布与定位。Western blot结果显示NS1基因编码蛋白获得表达,免疫荧光检测显示NS1蛋白主要存在于细胞核中。本研究为NS1蛋白功能和H5N1亚型AIV致病机制的研究奠定了基础。

To localized the nonstructure protein1（NS1） of human H5N1 serotype avian influenza virus（AIV） in infected cells,the NS1 gene was amplified by RT-PCR and cloned into eukaryotic expression plasmid of pXJ40-HA,the resultant recombinant plasmid was transfected into 293T cells,and the expressed NS1 was identified by western blot assay.The localization of NS1 protein in H1299 cells was detected by immunofluorescence assay（IFA）,and the results indicated that NS1 protein mainly localized in cell nucleus.This result would be benefited for further study of the function of NS1 protein and the mechanism of diseases induced by AIV serotype H5N1.