H5亚型禽流感病毒NS1基因在293T细胞中的瞬时表达与检测  

Transient expression and detection of nonstructural-1 gene of the subtype H5 avian influenza virus in 293T cells

作　　者：孙佳善[1,2] 文雪霞[1] 包红梅[1] 赵玉辉[1,2] 王云鹤[1] 熊永忠[1] 陈化兰[1] 王秀荣[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点开放实验室和农业部动物流感重点开放实验室,哈尔滨150001

出　　处：《黑龙江畜牧兽医》2013年第5期20-22,180,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金："十二五"农村领域国家科技课题项目(2012AA101303);哈尔滨市科技创新人才科研专项资金项目(RC2011XK002029);社会公益研究专项计划项目(200903055)

摘　　要：为了实现H5亚型禽流感病毒NS1基因的瞬时表达,试验利用PCR技术从NS1-T载体质粒上体外扩增NS1基因,将其克隆至pCAGGS载体,构建成重组质粒NS1-pCAGGS;提取NS1-pCAGGS质粒并与脂质体按照一定比例混合后转染293T细胞,用间接免疫荧光方法对瞬时表达细胞进行荧光信号反应。结果表明:禽流感NS1基因可以很好地在293T细胞中瞬时表达。To perform transient expression of nonstructural - 1 （ NS1 ） gene of the subtype H5 avian influenza virus, the NSI gene was amplified from NS1 - T vector plasmid in vitro and cloned into pCAGGS vector to construct a recombinant plasmid NS1 - pCAGGS. The NSI - pCAGGS plasmid was extracted, and then was mixed with liposome in accordance with a certain proportion to transfect the 293T cells. The indirect immu- nofluorescence assay was used to detect the fluorescence signal response from transient expression of cells. The result showed that the NSI gene of avian influenza virus could be transiently expressed well in 293T cells.

关 键 词：禽流感 NS1蛋白 瞬时表达 间接免疫荧光(IFA) 脂质体 

分 类 号：S852.659.5[农业科学—基础兽医学]