摘要
用大肠杆菌高效表达日本血吸虫中国大陆株 2 6k D GST基因 (Sj2 6)并观察表达产物诱导的免疫保护效果 .将 Sj2 6基因亚克隆至 p ET2 8b(+)中构建重组表达质粒 Sj2 6/ p ET,转化大肠杆菌 BL 2 1 (DE3) .Sj2 6在诱导条件下及未诱导条件下均获得高效表达 .表达产物 (r Sj2 6GST)以包含体形式表达 ,分子量为 2 8k D左右 ,能用 6× His亲和层析柱纯化 ,纯化产量为 55～ 60 mg/ L大肠杆菌培养物 .免疫印迹及 ELSA实验证实 r Sj2 6GST具有良好抗原性 .用 r Sj2 6GST不加佐剂直接背部皮下免疫昆明系小鼠 ,获得了 1 9.6% (P<0 .0 5)的减虫率及 31 .9% (P<0 .0 5)的成熟虫体减虫率 ;且免疫组检获的虫体中未成熟虫体的比例为 32 % (66/ 2 0 6) ,明显高于对照组的 1 9.6% (56/2 85) (P<0 .0 1 ) ,说明 r Sj2 6GST免疫不仅可诱导抗日本血吸虫的部分攻击感染作用 ,而且能抑制部分虫体的发育 .
The objective was to express the 26 kD glutathione S transferase(Sj26GST) gene of Chinese strain of Schistosoma japonicum in E.coli and observe the vaccination results of this expression products.The cDNA fragment encoding Sj26GST was cloned into the expression vector pET28b(+)and expressed in E.coli BL21(DE3) in conditions both of non induction and induction with IPTG.The recombinant protein (rSj26GST) was wholly insoluble in aquous solution and could be purified with 6×His chromatography column.Its molecular weight was about 28 kD.The yield was 55～60 mg/L E.coli culture.The immunological tests revealed that rSj26GST had good antigenicity.After vaccination in mice with rSj26GST,19.6% total worm reduction( P <0.05)and 31.9% matured worm reduction( P <0.05) were obtained.The ratio of immature worms in total worms harvested from vaccination group was 32%(66/206),significantly higher than that from control group(19.6%,56/285)( P <0.01).The results showed that vaccination with rSj26GST not only induced partial protection against Schistosoma japonicum infection,but also inhabited the development of Schistosoma japonicum in final host.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
2000年第6期759-763,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家科委863高技术计划资助!项目 (No.10 2 -0 7-0 4-0 3)
国家自然科学基金!资助项目 (39870 5 48)
