家蚕二分浓核病毒NS1蛋白单克隆抗体的制备及其特异性验证  

Preparation and Verification of Monoclonal Antibody Against Bom-byx mori Bidensovirus NS1 Protein

作　　者：胡朝阳[1] 刘伟[1] 邓炎春 唐培培 姚勤[1] 李国辉[1] Hu Zhaoyang;Liu Wei;Deng Yanchun;Tang Peipei;Yao Qin;Li Guohui(Institute of Life Sciences,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学生命科学研究院

出　　处：《蚕业科学》2018年第5期686-691,共6页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31570150,31270192)

摘　　要：家蚕二分浓核病毒(Bm BDV) NS1蛋白是一个与病毒复制相关的多功能蛋白。为获得NS1蛋白的单克隆抗体,设计并合成了12条NS1蛋白的抗原表位肽段,分别免疫BALB/c小鼠,取其脾细胞与SP2/0细胞融合,获得了18株能分泌抗体的阳性杂交瘤细胞。在大肠埃希菌中表达NS1作为抗原,通过Western blot对所获得的单克隆抗体进行验证。结果表明,由8/C278细胞株获得的单克隆抗体能特异识别重组的NS1蛋白,所用抗原表位序列为DIPPEEYRELRT。利用该抗体能检测到Bm BDV感染家蚕中肠组织中NS1蛋白的表达。该研究为深入探索NS1蛋白的功能打下了基础。The nonstructural protein NS1 of Bombyx mori bidensovirus( Bm BDV) is a multifunctional protein associated with virus replication. In order to obtain monoclonal antibody against NS1,twelve epitopes containing surface amino acid residues of NS1 protein were designed and synthesized. The synthetic polypeptides were used to immune female BALB/c mice respectively. After spleen cells of the immunized mice were used to fuse with SP2/0 cells,eighteen antibody-secreting positive hybridoma cell lines were obtained. The monoclonal antibodies were validated by Western blot using recombinant NS1 expressed in Escherichia coli as antigen. Results showed that the monoclonal antibody secreted by 8/C278 cell line could specifically recognize NS1 protein,the sequence of epitope corresponding to this antibody is DIPPEEYRELRT. NS1 protein could be detected in midgut of Bm BDV-infected silkworm by Western blot using 8/C278 as first antibody. This study will be beneficial for further functional study of NS1 protein of Bm BDV.

关 键 词：家蚕二分浓核病毒 NS1蛋白 单克隆抗体 抗原表位 

分 类 号：S852.659.2[农业科学—基础兽医学]