中国兽医科学2026，Vol.56Issue(1)：40-49,10.DOI:10.16656/j.issn.1673-4696.2025.0242
猪囊尾蚴丝氨酸蛋白酶抑制剂B6和4848优势抗原表位的分析与鉴定
Analysis and identification of dominant antigenic epitopes of serine protease inhibitors B6 and 4848 from Cysticercus cellulosae
摘要
Abstract
Aiming at the problems of insufficient exposure of Cysticercus cellulosae serine protease inhibitors B6(Ts-serpin B6)and 4848(Ts-serpin 4848)antigenic epitopes and lack of high specificity,it is proposed to enhance the antigenic specificity by screening key antigenic epitopes.The sequences of Ts-serpin B6 and Ts-serpin 4848 genes were firstly analyzed by bioinformatics epitope analysis to screen out the dominant antigenic epitopes.After designing specific primers,the target gene fragments were amplified respectively by PCR.The purified PCR products were ligated into pET-28a(+)vector and then transformed into Escherichia coli DH5α receptor cells to construct segmented prokaryotic expression vectors for pET28a(+)-Ts-serpin B6 and pET28a(+)-Ts-serpin 4848,which were transformed into E.coli BL21(DE3)to induce express ion,and the target proteins were purified and then identified by SDS-PAGE and analyzed by Western-blot.Six recombinant target proteins,Ts-serpin B6(a),Ts-serpin B6(b),Ts-serpin B6(c),and Ts-serpin 4848(a),Ts-serpin 4848(b),Ts-serpin 4848(c),which existed in E.coli mainly in the form of inclusion bodies,were successfully expressed and purified.The results of Western-blot and ELISA showed that the recombinant proteins Ts-serpin B6(a)and Ts-serpin 4848(a)had stronger reactogenicity and specificity compared with other segmented recombinant proteins.This study clarified that Ts-serpin B6(a)and Ts-serpin 4848(a)are the dominant epitopes of the antigens,which provides a basis for the development of diagnostic reagents specific for C.cellulosae.关键词
猪囊尾蚴/丝氨酸蛋白酶抑制剂/优势抗原表位/鉴定表达Key words
Cysticercus cellulosae/serine protease inhibitor/dominant antigenic epitope/iden-tification of expression分类
农业科技引用本文复制引用
段蕊,王思睿,赵昱源,王梦奇,陆沪江,何沛霞,孙树民..猪囊尾蚴丝氨酸蛋白酶抑制剂B6和4848优势抗原表位的分析与鉴定[J].中国兽医科学,2026,56(1):40-49,10.基金项目
国家自然科学基金项目(31960707,32160842,32460893) （31960707,32160842,32460893）
内蒙古自治区科技计划项目(2022YFDZ0049) （2022YFDZ0049）
云南省科技厅科技计划项目(202504BI090003) （202504BI090003）
内蒙古自治区直属高校基本科研业务费项目(GXKY22042) （GXKY22042）
云南省重大科技专项(202402AE090074) （202402AE090074）
建设面向南亚东南亚科技创新中心专项(202403AP140033) （202403AP140033）
