现代畜牧科技Issue(8)：22-26,5.DOI:10.19369/j.cnki.2095-9737.2024.08.005
PRRSV GP2-GP4融合蛋白原核表达及多克隆抗体的制备
Prokaryotic Expression of PRRSV GP2-GP4 Protein and Preparation of Polyclonal Antibody
摘要
Abstract
Porcine reproductive and respiratory syndrome virus(PRRSV)is one of the important pathogens that harm the development of pig industry.Since the structural proteins GP2 and GP4 can bind to the CD163 receptor of target cells and can participate in the virus neutralization reaction,the study for the GP2 and GP4 protein is significant for PRRSV vaccine development.In this study,GP2 and GP4 genes were linked by flexible peptides,cloned into the prokaryotic expression vector pET32a,and the GP2-GP4 fusion protein was successfully expressed through the expression system of E.coli.The target protein was expressed in the form of an inclusion bodies,and high purity GP2-GP4 fusion protein was obtained after purification and refolding.The polyclonal antibodies were prepared by immunizing rabbits.SDS-PAGE results showed that the size of the target protein was about 55 kDa,which was consistent with the expectation.ELISA revealed a GP2-GP4 antibody titer of 1∶64000.Western blot and IFA results showed that polyclonal antibody could react specifically with GP2-GP4 protein with good reactivity.In summary,the fusion expression of GP2-GP4 proteins was obtained and the polyclonal antibody was prepared with good specificity,which laid a foundation for the investigation of the function of PRRSV GP2-GP4 proteins and subsequent vaccine research.关键词
猪繁殖与呼吸综合征/糖蛋白/原核表达Key words
porcine reproductive and respiratory syndrome/glycoprotein/prokaryotic expression分类
农业科技引用本文复制引用
宁慧敏,彭娜娜,陈玉豪,李欣颖,范杰,董伟..PRRSV GP2-GP4融合蛋白原核表达及多克隆抗体的制备[J].现代畜牧科技,2024,(8):22-26,5.基金项目
云南省重大科技专项计划(202202AE090032) （202202AE090032）
