猪流行性腹泻病毒变异毒株中和表位区S10原核表达及多克隆抗体的制备  

Prokaryotic Expression of Neutralizing Epitope Regin S10 and Preparation of Polyclonal Antibody Against Porcine Epidemic Diarrhea Virus Variant

作　　者：王美娇 杨丹[1] 赵飞宇 李春秋[1] 朱庆贺 邢晓旭 孙东波[1] Wang Meijiao;Yang Dan;Zhao Feiyu;Li Chunqiu;Zhu Qinghe;Xing Xiaoxu;Sun Dongbo(College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Daqing163319)

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319

出　　处：《黑龙江八一农垦大学学报》2024年第6期27-35,共9页journal of heilongjiang bayi agricultural university

基　　金：黑龙江省重点研发计划指导项目(GZ20220029)。

摘　　要：试验对猪流行性腹泻病毒(PEDV)变异毒株S1的S10表位区进行表达,并制备抗S10蛋白多克隆抗体。将S10区基因连接至原核表达载体pET-32a(+),经大肠杆菌原核表达系统成功表达pET-32a-S10重组蛋白,重组蛋白分子量约为41 kDa,利用Ni柱进行亲和纯化后浓度为2.01mg·mL^(-1)。纯化的重组蛋白经Western blot鉴定后作为免疫原免疫BALB/c小鼠,并收集小鼠血清,利用间接ELISA方法检测血清中抗体效价可达到1∶12800,该抗体可以结合S10重组蛋白和天然PEDV HM2017毒株。研究成功制备了良好免疫原性的PEDV S10蛋白多克隆抗体,为进一步研究S1蛋白及PEDV诊断方法的建立奠定基础。The S10 epitope region of porcine epidemic diarrhea virus(PEDV)variant S1 was expressed and polyclonal antibody against S10 protein was prepared.The S10 region gene was linked to the prokaryotic expression vector pET-32a(+),and the recombinant protein PET-32A-S10 was successfully expressed by the prokaryotic expression system of Escoli.The molecular weight of the recombinant protein was about 41 kDa,and the concentration of the recombinant protein was 2.01 mg·mL^(-1) after affinity purification by Ni column.After Western blot identification,the purified recombinant protein was used as an immunogenic agent to immunize BALB/c mice,and mouse serum was collected.The titer of antibody in serum was detected by indirect ELISA method,which could reach 1∶12800,and the antibody could bind S10 recombinant protein and natural PEDV HM2017 strain.The study successfully prepared PEDV S10 protein polyclonal antibody with good immunogenicity,which laid a foundation for further study of S1 protein and establishment of PEDV diagnosis method.

关 键 词：猪流行性腹泻病毒 S1蛋白 中和表位区S10 多克隆抗体 

分 类 号：S858.28[农业科学—临床兽医学]