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目的 鉴定纯化的重组日本血吸虫线粒体相关融合蛋白 (r Sj3 3 8/2 6GST)的免疫原性。方法 对已构建的阳性表达菌 p GEX-6p-1/Sj3 3 8进行大量诱导表达 ,并对纯化蛋白进行活性分析。将粗提包涵体蛋白、复性包涵体蛋白、经分子筛纯化并复性的蛋白分别免疫新西兰家兔 2只。用 West-ern blot及 Dot-EL ISA方法检测特异性抗体的免疫原性及各组抗体水平。结果 用分子筛纯化的融合蛋白 r Sj3 3 8/2 6GST经 SDS-PAGE电泳鉴定显示达电泳纯 ;Western blot显示纯化蛋白能够被日本血吸虫重感染兔血清及 r Sj3 3 8/2 6GST免疫兔血清识别。Dot-ELISA法检测结果表明 ,经分子筛纯化并复性的 r Sj3 3 8/2 6GST融合蛋白免疫兔血清中特异性抗体的滴度最高 ,为 1∶ 5 12 0 0。结论 纯化后的融合蛋白 r Sj3 3 8/2 6GST具有较高的纯度及较强的免疫活性 ,可望作为日本血吸虫病疫苗候选分子 ,值得进一步深入研究
Objective To identify the immunogenicity of purified recombinant Schistosoma japonicum mitochondria-associated fusion protein (r Sj3 3 8/2 6GST). Methods The constructed positive expression strain pGEX-6p-1 / Sj3 38 was induced in large quantities and the activity of the purified protein was analyzed. Crude inclusion protein, refolding inclusion protein, molecular sieve purification and renaturation of proteins were immunized New Zealand rabbits 2. The immunogenicity and antibody levels of specific antibodies were detected by West-ern blot and Dot-ELISA method. Results The fusion protein r Sj3 3 8/2 6GST purified by molecular sieve was electrophoresed on SDS-PAGE. Western blot showed that the purified protein could be recognized by rabbit sera of Schistosoma japonicum and sera of r Sj3 3 8/2 6 GST . The results of Dot-ELISA showed that the titer of specific antibody in rabbit serum immunized with rSj3 3 8/2 6GST fusion protein purified by molecular sieve and refolded was the highest, which was 1:5 1200. Conclusion The purified fusion protein r Sj3 3 8/2 6GST has high purity and strong immunogenicity and is expected to be a candidate vaccine candidate for schistosomiasis japonica, which deserves further study