中国动物传染病学报2024，Vol.32Issue(6)：25-35,11.
融合蛋白EMP-VP2在杆状病毒表达系统中的表达与免疫原性鉴定
Baculovirus Expression and Immunogenicity of Fusion Protein Emp-VP2 of Japanese Encephalitis Virus and Porcine Parvovirus
摘要
Abstract
Japanese encephalitis virus(JEV)and Porcine parvovirus(PPV)cause reproductive disorders in pigs,which seriously affect the economic benefits of the pig industry.In this study,multiple B cell and T cell antigenic epitopes on the E gene of Japanese encephalitis virus(JEV)were fused with the VP2 gene of porcine parvovirus(PPV)to construct the pFastBac Dual-EMP-VP2 vector.After transforming into DH10Bac competent cells,the rBacmid EMP-VP2 recombinant bacmid was successfully constructed through blue-white screening.The recombinant baculovirus was then successfully rescued in Sf9 insect cells,achieving efficient expression of EMP-VP2.SDS-PAGE and Western blot showed that the molecular weight of the expressed product was in line with expectations.The expressed recombinant EMP-VP2 protein reacted with JEV and PPV positive serum samples,indicating its good immunoreactivity.Subsequently,the expressed protein was purified through affinity chromatography and used to immunize mice and guinea pigs.These animals produced specific humoral immune responses to JEV and PPV,which proved that the EMP-VP2 protein had immunogenicity,laid the foundation for development of a binary subunit vaccine for prevention and control of JEV and PPV.关键词
日本脑炎病毒/猪细小病毒/杆状病毒表达系统/免疫反应性/免疫原性Key words
Japanese encephalitis virus/Porcine parvovirus/baculovirus expression system/immunoreactivity/immunogenicity分类
农业科技引用本文复制引用
张欣雅,马志永,杨扬,田羽彤,李宗杰,李蓓蓓,刘珂,邵东华,邱亚峰,魏建超..融合蛋白EMP-VP2在杆状病毒表达系统中的表达与免疫原性鉴定[J].中国动物传染病学报,2024,32(6):25-35,11.基金项目
国家重点研发计划项目(2022YFD1800801) （2022YFD1800801）
上海市科委农业领域项目(22N41900400) （22N41900400）
