ClassⅠ新城疫病毒核衣壳蛋白在体外细胞感染过程中表达定位的检测  

Dynamic distribution of NP protein in class INewcastle disease virus infected DF1 cells

作　　者：孙英杰[1] 陈鸿军[1] 詹媛[1] 仇旭升[1] 于洋[1] 于圣青[1] 丁铲[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国预防兽医学报》2011年第2期85-88,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30871883);2007年中央公益性研究院所基础业务专项(2007JB09)

摘　　要：为分析核衣壳蛋白(NP)在新城疫病毒(NDV)感染真核细胞过程中细胞内定位的情况,本研究以NDV Class I强毒株9a5b作为免疫原制备4株NP特异性单克隆抗体(MAb),并将病毒株9a5b按5 MOI感染量接种DF1单层细胞,当接种后1 h～3 h时,NP有极少量表达并逐量增加,主要呈点状分布于细胞浆内;当接种后4 h～20 h时,NP聚集于细胞浆中,聚集数量与感染时间成正比;感染24 h后,细胞核碎裂,NP散在分布于细胞浆中。表明NP作为立早期蛋白,始终在胞浆中进行复制;在病毒感染初期,NP介导核糖核蛋白复合物发挥转录和翻译功能。本研究为深入开展NP与病毒mRNA转录、复制和包装的相互关系机制的研究奠定了基础。To prepare monoclonal antibodies（MAb） against Nucleoprotein（NP） of class I Newcastle disease virus（NDV）,a virulent strain,designated as 9a5b,was served as the antigen.A panel of 4 hybridoma cell lines was gained to prepare MAb specific to NP protein.The MAbs were used to analyze the dynamic distribution of NP protein in DF1 cells infected by NDV.The DF1 monolayer cells were infected with NDV 9a5b strain at a dose of 5 M.O.I.A small quantity of NP protein was expressed and diffusely distributed in cytoplasm at 1 to 3 hours post infection（pi）.NP protein accumulated in cytoplasm and the fluorescence intensity gradually increased at 20 hours pi.With the nuclei fused together,NP protein also scattered in the cytoplasm after 20 hours pi.The relationship between NP protein and RNA assembly or virus replication of NDV will be investigated in the future.

关 键 词：新城疫病毒 ClassⅠ 核蛋白 细胞内定位 

分 类 号：S852.65[农业科学—基础兽医学]