日本乙型脑炎病毒SA14-14-2株NS1-Flag蛋白在293T细胞中的表达  

Eukaryotic Expression of NS1-Flag Protein of Japanese Encephalitis Virus SA14-14-2 in 293T Cells

作　　者：臧富玉[1] 亓文宝[1] 陈孝明[1] 刘国乾[1] 李红梅[1] 廖明[1] 

机构地区：[1]华南农业大学兽医学院农业部动物疫病防控重点开放实验室,广东广州510642

出　　处：《中国畜牧兽医》2011年第9期40-43,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(30800827);广东省科技计划项目(2009B030803049;2010B020307009);兽医生物技术国家重点实验室开放基金(SKLVBF201002);广东省自然科学基金(06105471)

摘　　要：通过RT-PCR方法扩增出日本乙型脑炎病毒SA14-14-2株的NS1基因,利用HindⅢ和HpaⅠ双酶切克隆到带有Flag标签的pcDNA3.1载体上,重组质粒命名为pcDNA3.1-NS1-Flag。脂质体法转染293T细胞后,经间接免疫荧光(IFA)和化学荧光Western blotting检测,证明带有Flag标签的NS1基因在293T细胞中得到高效表达。RT-PCR was performed on the total RNA extracted from Japanese encephalitis virus（JEV）SA14-14-2 to obtain the cDNA of NS1,the cDNA was edonuclease digestion by Hind Ⅲ/HpaⅠand cloned into a vector pcDNA3.1-Flag.The recombinant plasmid was named plasmid pcDNA3.1-NS1-Flag,identified correctly by DNA sequencing and restriction enzyme analysis.The plasmid pcDNA3.1-NS1-Flag was transfected into 293T cell by means of liposome and its expression was examined by immunofluorescence assay and Western blotting,the results showed that NS1 protein could express in 293T cell.

关 键 词：日本乙型脑炎 NS1-Flag融合蛋白 真核表达 293T细胞 

分 类 号：Q78[生物学—分子生物学]