禽流感病毒NS1蛋白在杆状病毒表达系统中的表达  

Expression of Avian Influenza Virus NS1 Protein in Baculovirus Expression Vector System

作　　者：李东卫[1,2] 郭莹莹[1] 刘在斯[1] 王世达[1,2] 沈楠[1] 文辉强[1] 焦同路[1] 王靖飞[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所动物疫病诊断中心农业部兽用药物与兽医生物技术重点实验室,黑龙江哈尔滨150001 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《动物医学进展》2012年第6期27-31,共5页Progress In Veterinary Medicine

基　　金：中国农业科学院哈尔滨兽医研究所院所长基金(SKLVBP201016)

摘　　要：为获得纯化的禽流感病毒(AIV)的NS1蛋白,将A/chicken/Guangxi/10/99(H9N2)(CK/GX/10/99)NS1全长核苷酸克隆至杆状病毒转移载体pFastBacHTA中,构建了重组转移载体pFastBacHTA-NS1,然后转化入DH10Bac感受态细胞中制备重组杆粒,在脂质体介导下转染sf9昆虫细胞,获得表达NS1基因的重组杆状病毒。用重组杆状病毒转染昆虫细胞sf9进行NS1蛋白的表达,通过SDS-PAGE电泳、Western blot、间接免疫荧光(IFA)对表达的目标蛋白进行分析。结果表明,构建了含NS1基因的重组杆状病毒,分子质量约30ku的重组蛋白得到了表达,为NS1的相关功能研究奠定了基础。In order to obtain purified avian influenza virus NS1 protein, NSlgene of A/chicken/Guangxi/ 10/99（H9N2） （CK/GX/10/99） was cloned into baculovirus transfer vector pFastBacHTA and constructed a recombinant transfer vector pFastBacHTA-NS1. Plasmid pFastBacHTA-NS1 was then transformed into DH10Bac competent cells to obtain recombinant bacmid after the recombinant Bacmid DNA was isolated and transfected into Sf9 insect cells to generate a recombinant NS1 expressing baculovirus which was transfected into Sf9 insect cells to express NS1 protein. The expressed NS1 protein was tested using SDSPAGE, Western blot and indirect immunofluorescence assay（IFA）. The results showed that the recombinant baeulovirus containing NS1 gene was constructed successfully and the recombinant protein was successfully expressed with a relative molecular of 30 ku. This work provides the basic material for NS1 related function research.

关 键 词：禽流感病毒 NS1蛋白 杆状病毒表达载体系统 蛋白纯化 

分 类 号：S852.657[农业科学—基础兽医学]