斜纹夜蛾核型多角体病毒单克隆抗体的制备和分析  

Preparation and Characterization of Monoclonal Antibodies Against Spodoptera litura Nucleopolyhedrovirus

作　　者：张亚力[1] 钟江[1] 苏德明[1] 庞义[2] 

机构地区：[1]复旦大学生命科学学院微生物学与微生物工程学系,上海200433 [2]中山大学生物防治国家重点实验室,广州510257

出　　处：《复旦学报（自然科学版）》2001年第5期504-508,共5页Journal of Fudan University：Natural Science

基　　金：国家自然科学基金重点项目(38730030);教育部重点实验室访问学者基金资助课题

摘　　要：用纯化的斜纹夜蛾核型多角体病毒（Ｓｐｏｄｏｐｔｅｒａ　ｌｉｔｕｒａｎｕｃｌｅｏ　ｐｏｌｙｈｅｄｒｏｖｉｒｕｓ）的病毒核衣壳免疫小鼠，取脾脏细胞与骨髓瘤细胞融合，经筛选得到５株　稳定分泌单克隆抗体的杂交瘤细胞株．它们所分泌的单抗分属ＩｇＧ１，ＩｇＧ２ａ，ＩｇＧ　２ｂ和ＩｇＧ３等４种抗体亚类，其中１株单抗识别分子质量为４１　ｋｕ的病毒蛋白，其　余４株单抗均识别３１　ｋｕ的病毒蛋白．胰蛋白酶部分酶解分析发现，３１　ｋｕ蛋白的４株单抗分别　识别至少３个不同的抗原决定簇．所有５株单抗均不与其他４种核型多角体病毒发生交叉反应　．利用所制备的单抗进行了病毒在虫体中增殖动态的检测．这些单克隆抗体可用来深入研究　病毒的流行规律和复制机制．Five hybridoma cell lines that secreted monoclonal antibodies against the armyworm Spodoptera litura were established.These mono clonal antibodies belonged to four different IgG subclasses of IgG 1 , IgG 2a , IgG 2b , and IgG 3 . Western blot analysis revealed that Mab A7D 6 specifically reacted with the 41 ku viral peptide, while the other four Mabs reacted with the 31 ku viral peptide.Trypsin partial digestion experiment demonstrated that these four Mabs recognized at least three different epitopes. These Mabs did not react with four other nucleopolyhedroviruses, whereas the polyclonal antibodies against SpltNPV cross reacted with them.The mon oclonal antibodies were currently used to investigate the dynamics of virus multiplication in infected larvae and in the field.

关 键 词：斜纹夜蛾 核型多角体病毒 杂交瘤 单克隆抗体 杆状病毒科 生物防治 病毒复制 复制机制 

分 类 号：Q939.4[生物学—微生物学] S476.13[农业科学—农业昆虫与害虫防治]