农业生物技术学报2026，Vol.34Issue(3)：598-609,12.DOI:10.3969/j.issn.1674-7968.2026.03.013
口蹄疫多表位重组蛋白在大肠杆菌中的可溶性表达及其免疫效果评价
Soluble Expression of Foot-and-mouth Disease Multi-epitope Recombinant Protein in Escherichia coli and Evaluation of Its Immunization Effect
摘要
Abstract
Foot-and-mouth disease(FMD),caused by the Foot-and-mouth disease virus(FMDV),is a highly contagious disease among cloven-hoofed animals,posing a serious threat to the development of the livestock industry.In order to develop a novel epitope vaccine for FMD,this study,based on the gene sequence of the prevalent swine(Sus scrofa)O-type FMDV strain(O/MYA98/BY/2010),constructed a recombinant expression plasmid pColdⅡ-7B2T containing 7 B-cell epitopes and 2 T-cell epitopes.The recombinant plasmid was co-expressed with the molecular chaperone TF16 in Escherichia coli,followed by a series of analytical procedures including affinity purification,SDS-PAGE,and Western blot analysis.The recombinant protein was then used to immunise BALB/c mice(Mus musculus),after which its humoral and cellular immune responses were evaluated.The results demonstrated that the 7B2T recombinant protein,prepared using the molecular chaperone TF16,was expressed in a soluble form with a molecular weight of approximately 26 kD and could induce a specific immune response with FMDV-positive serum.Furthermore,the 7B2T protein was found to induce significant specific IgG production in the host,with an average neutralising antibody titer of 1∶96.In addition,the 7B2T protein had been observed to induce robust cellular immune responses in murine models,promoting the differentiation of CD4+and CD8+T lymphocyte subsets within the spleen.Lymphocyte proliferation assays and enzyme-linked immunospot assays(ELISPOTS)revealed that the splenic lymphocyte proliferation levels and interferon-γ(IFN-γ)spot counts in the 7B2T protein-treated group were extremely significantly higher than those in the inactivated vaccine(IV)group and PBS group(P＜0.01),while the IL-4 spot counts were comparable to those of the IV group.In summary,the present study demonstrated that the soluble multi-epitope 7B2T recombinant protein,prepared using the molecular chaperone TF16,was capable of inducing both humoral and cellular immune responses in mice.This study provides experimental ideas and data references for the preparation of FMD multi-epitope recombinant proteins and the development of multi-epitope vaccines.关键词
口蹄疫病毒/原核表达/分子伴侣/表位疫苗/免疫原性Key words
Foot-and-mouth disease virus/Prokaryotic expression/Molecular chaperones/Epitope vaccine/Immunogenicity分类
农业科技引用本文复制引用
蒋成辉,茹毅,郝荣增,李亚军,赵陇和,杨洋,卢炳州,郑海学,曾巧英..口蹄疫多表位重组蛋白在大肠杆菌中的可溶性表达及其免疫效果评价[J].农业生物技术学报,2026,34(3):598-609,12.基金项目
甘肃省自然科学基金重点项目(23YFNA0011) （23YFNA0011）
甘肃省研产融合科技攻关赋能计划(25FNNA002) （25FNNA002）
国家自然科学基金(32372990) （32372990）
甘肃省自然科学基金(23JRRA549) （23JRRA549）
中国农业科学院兰州兽医研究所所级基本科研业务费(1610312021013) （1610312021013）
甘肃省科技重大专项计划(23ZDNA007) （23ZDNA007）
国家生猪技术创新中心先导科技项目(NCTIP-XD/C03) （NCTIP-XD/C03）
国家生猪产业技术体系(CARS-35) （CARS-35）
中国农业科学院兰州兽医研究所所级基本科研业务费(1610312022009) （1610312022009）
