黑龙江八一农垦大学学报2024，Vol.36Issue(6)：27-35,9.DOI:10.3969/j.issn.1002-2090.2024.06.005
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
摘要
Abstract
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∶12 800,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Key words
porcine epidemic diarrhea virus/S1 protein/neutralizing epitope region S10/polyclonal antibody分类
农业科技引用本文复制引用
王美娇,杨丹,赵飞宇,李春秋,朱庆贺,邢晓旭,孙东波..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[J].黑龙江八一农垦大学学报,2024,36(6):27-35,9.基金项目
黑龙江省重点研发计划指导项目(GZ20220029). （GZ20220029）
