中国兽医科学2025，Vol.55Issue(11)：1509-1516,8.DOI:10.16656/j.issn.1673-4696.2025.0214
非洲猪瘟病毒pE199L蛋白在大肠杆菌中的表达及免疫原性分析
Prokaryotic expression and immunogenicity evaluation of African swine fever virus pE199L protein
摘要
Abstract
This study aimed to express and purify the African swine fever virus(ASFV)envelope protein pE199L using an Escherichia coli expression system and evaluate its immunogenicity.The codon optimized gene encoding pE199L was cloned into the pET-32a vector,and the recombinant plasmid was expressed in E.coli.The target protein was purified by affinity chromatography and verified for specificity by Western-blot.Mice were immunized with the purified protein to produce polyclonal antibodies.The antibody titer and specificity were respectively assessed by indirect ELISA and indirect immunofluorescence assay,and the antiviral activity of the immune serum was evaluated through an ASFV replication inhibition assay.The results showed successful production of the pE199L protein,with a serum antibody titer reaching 1∶128 000 on day 28 post-immunization.The serum specifically recognized ASFV-infected bone marrow-derived macrophages.Flow cytometry revealed significant activation of CD4+and CD8+T lymphocytes in the spleen,accompanied by elevated levels of IFN-γ and IL-4 in the serum.The replication inhibition assay demonstrated that antibodies induced by pE199L effectively suppressed ASFV replication.In conclusion,pE199L exhibits strong immunogenicity and represents a promising candidate antigen for the development of ASFV subunit vaccines.关键词
非洲猪瘟/pE199L蛋白/原核表达/免疫原性Key words
African swine fever/pE199L protein/prokaryotic expression/immunogenicity分类
畜牧业引用本文复制引用
张亮,张越,安芳兰,杨进才..非洲猪瘟病毒pE199L蛋白在大肠杆菌中的表达及免疫原性分析[J].中国兽医科学,2025,55(11):1509-1516,8.基金项目
非洲猪瘟亚单位疫苗产业化生产技术体系创建与推广应用项目(23ZDKA0002) （23ZDKA0002）
甘肃省自然科学基金重点项目(23YFNA0011) （23YFNA0011）
中央级公益性科研院所基本科研业务费专项(CAAS-ZDRW202409) （CAAS-ZDRW202409）
甘肃省科技重大专项(23ZDNA007) （23ZDNA007）
