论文部分内容阅读
目的克隆、表达日本血吸虫肌钙蛋白T(SjTnT)编码cDNA,评估重组抗原抗血吸虫感染的免疫保护效果。方法利用PCR技术扩增SjTnT基因,构建重组表达质粒pET28a(+)-SjTnT并诱导表达,用重组蛋白rSjTnT免疫BALB/c小鼠制备免疫血清。利用免疫印迹试验(Western blotting)和酶联免疫吸附试验(ELISA)检测rSjTnT诱导的免疫反应。采用rSjTnT免疫小鼠,评估其诱导的免疫保护效果。结果成功克隆、表达了日本血吸虫SjTnT基因。Western blotting显示rSjTnT能被该重组蛋白免疫小鼠血清识别,具有良好的免疫原性。rSjTnT能诱导小鼠产生高水平的特异性IgG抗体。动物免疫保护试验表明,rSjTnT能诱导小鼠产生33.89%的减虫率以及43.94%的肝脏减卵率。结论制备的rSjTnT在小鼠体内能诱导产生部分抗血吸虫感染的免疫保护效果,为评估SjTnT的生物学功能奠定了基础。
Objective To clone and express SjTnT cDNA of Schistosoma japonicum, and to evaluate the immunoprotective effect of recombinant antigens against schistosoma infection. Methods The SjTnT gene was amplified by PCR, and the recombinant plasmid pET28a (+) - SjTnT was constructed and induced. The recombinant protein rSjTnT was used to immunize BALB / c mice to prepare the immune serum. The immunological reaction induced by rSjTnT was detected by Western blotting and enzyme-linked immunosorbent assay (ELISA). Mice were immunized with rSjTnT to evaluate their induced immune protective effect. Results The gene SjTnT was successfully cloned and expressed in Schistosoma japonicum. Western blotting showed that rSjTnT could be recognized by the serum of mice immunized with the recombinant protein and had good immunogenicity. rSjTnT induced mice to produce high levels of specific IgG antibodies. Animal immune protection test showed that rSjTnT can induce mice to produce 33.89% worm reduction rate and 43.94% of the liver reduction rate. Conclusion The rSjTnT prepared in mice can induce partial immunosuppressive effect of anti-schistosome infection, which lays the foundation for evaluating the biological function of SjTnT.