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目的 在大肠杆菌中高效表达日本血吸虫 (Schistosomajaponicum Sj)Mf2蛋白 ,并对表达产物进行免疫保护效果测定。 方法 将SjMf2基因亚克隆至pCEX -5X -3原核表达载体 ,以GST融合蛋白的形式在ER2 688中表达 ,表达产物免疫小鼠 ,免疫用抗原剂量为 5 0 μg 次 鼠 ,对照组分别注射FCA或PBS ,在 0、2、6周共免疫 3次。第 3次免疫后 2周进行攻击感染 ,42d后杀鼠 ,观察减虫和减卵效果。 结果 在IPTG诱导下 ,表达载体中的SjGST基因与重组的SjMf2基因在大肠杆菌中得以高效表达 ,形成了SjGST -Mf2融合蛋白 ,用这种融合蛋白免疫小鼠 ,诱导产生了2 7 75 %的减虫率和 45 7%的每雌肝组织虫卵减少率 (LEPF)。 结论 SjMf2基因亚克隆至pGEX -5X -3载体后可在大肠杆菌中高效表达 ,表达产物能诱导小鼠产生一定程度的抗日本血吸虫保护性免疫力。
Objective To express Mf2 protein of Schistosoma japonicum Sj efficiently in Escherichia coli and determine its immunoprotective effect. Methods SjMf2 gene was subcloned into pCEX-5X-3 prokaryotic expression vector and expressed in ER2 688 as a GST fusion protein. The immunized mice were immunized with the antigen dose of 50 μg and the control group were injected with FCA Or PBS, co-immunized 3 times at 0, 2, 6 weeks. Two weeks after the third immunization, the challenge was carried out. After 42 days, the mice were killed and the effects of de-worming and egg reduction were observed. Results Under the induction of IPTG, the SjGST gene in the expression vector and the recombinant SjMf2 gene were highly expressed in E. coli and the SjGST-Mf2 fusion protein was formed. Immunization of mice with this fusion protein resulted in the production of 27 75% Worm reduction rate and 45 7% reduction in egg loss per female liver tissue (LEPF). Conclusion The SjMf2 gene was subcloned into pGEX - 5X -3 vector and expressed efficiently in E. coli. The expressed product of SjMf2 gene could induce some degree of protective immunity against Schistosoma japonicum in mice.