Asia 1型口蹄疫病毒VP1基因的表达鉴定及其多抗制备  

Expression of VP1 gene of foot-and-mouth disease virus serotype Asia 1 and antiserum preparation

作　　者：朱远茂[1,2] 任宪刚 史鸿飞[2] 高欲燃[1,2] 于作[2] 冯军科[2] 相文华[1,2] 薛飞[2] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨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实验室/大动物传染病研究室,黑龙江哈尔滨150001 [3]哈药集团生物疫苗有限公司,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2010年第4期285-288,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家863计划(2006AA10A204)

摘　　要：本研究旨在表达口蹄疫病毒(FMDV)的VP1全基因并制备特异性的多克隆抗体。利用PCR方法扩增Asia 1 IND 49197株VP1全基因,将其克隆至原核表达载体pET-30a(+)中,在大肠杆菌BL21中进行表达。SDS-PAGE结果显示表达产物分子量约为31.6ku,以包涵体的形式存在。通过Ni-NTA Purification System纯化后进行western blot和间接ELISA分析,结果显示重组蛋白能够被FMD阳性血清识别,具有良好的反应性。将纯化的重组蛋白免疫新西兰白兔制备多克隆抗体,ELISA测定抗体效价为1∶20480,病毒中和试验测定抗体效价为1∶64。本研究所表达的VP1蛋白可用于开发检测Asia1口蹄疫抗体的诊断试剂,所制备的多克隆抗体为进一步研究VP1的结构、功能以及抗原表位的鉴定提供了条件。In this study,the VP1 gene of foot and mouth disease virus（FMDV） was amplified from Asia 1 IND 4/2004 strain by PCR and cloned into the pET-30a（＋） for expression in E. coli BL21. The expressed protein had a molecular weight of approximately 31.6 ku,and could react specifically to FMD positive serum in indirect ELISA and western blot analysis. Polyclonal antibodies to the VP1 protein was prepared by inoculating rabbits with purified recombinant VP1 protein followed by boosting 3 times. Anti-FMDV-VP1 serum were generated with antibody titres of 1∶20 480 in ELISA and 1∶64 in virus neutralization test. The recombinant protein obtained in this study could be used for detection of FMDV Asia 1 antibodies,and the polyclonal antibodies could be used to further study the structure,function and epitope mapping of FMDV serotype Asia 1 VP1 gene.

关 键 词：ASIA I口蹄疫病毒 VP1基因 多克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]