四川大学学报（自然科学版）2025，Vol.62Issue(2)：494-500,7.DOI:10.19907/j.0490-6756.240260
重组球菌和球虫多表位抗原的表达及特性研究
Expression and characterization of recombinant multi-epitope antigens of cocci and coccidia
摘要
Abstract
This study aimed to develop a combined multi-epitope vaccine targeting both Streptococcus bovis and Bovine coccidia.Using bioinformatics approaches,dominant T-cell and B-cell epitope gene sequences from the two pathogens were identified and tandemly linked,followed by the incorporation of cholera toxin B subunit(CTB)as an internal molecular adjuvant to design the recombinant antigen CARR(CTB-AMA1-RodA-RodA).Following codon optimization,the CARR gene was cloned into the pET28a(+)vector and transformed into E.coli BL21(DE3),thereby establishing a stable expression strain.SDS-PAGE analysis confirmed the recombinant strain expressed a target protein(approximately 90 kDa),and Western blot con-firmed its specific binding to both anti-CTB and anti-Streptococcus bovis antibodies,with a plasmid retention rate above 80%after 20 generations.The purified CARR protein was used to immunize laying hens,and ELISA results demonstrated that the specific IgY antibody titer reached 1:12,800 after 7 weeks.In vitro neu-tralization assays confirmed that the antibodies significantly inhibited the growth of S.bovis,indicating potent functional neutralizing activity.These results suggest that the CARR recombinant protein can effectively and rapidly elicit a robust humoral immune response in the host.This study provides a promising candidate anti-gen and technical foundation for the development of a combined vaccine against S.bovis and E.bovis infec-tions.关键词
牛链球菌/牛艾美尔球虫/重组多表位抗原/表达特性/中和抗体Key words
Streptococcus bovis/Eimeria bovis/Recombinant multi-epitope antigen/Expression characteris-tics/Neutralizing antibodies分类
医药卫生引用本文复制引用
符立发,别明江,王保宁,梁鸽,黄家艳,唐智慧,赵仕杰,张焱荣,郑璨璘,索朗斯珠,曹随忠..重组球菌和球虫多表位抗原的表达及特性研究[J].四川大学学报（自然科学版）,2025,62(2):494-500,7.基金项目
西藏自治区科技厅重点研发计划项目(XZ202201ZY0009N) （XZ202201ZY0009N）
四川省科技厅项目(2023YFQ0051) （2023YFQ0051）
