弓形虫棒状体蛋白ROP38的原核表达与鉴定

目的原核表达弓形虫棒状体蛋白ROP38，并对重组蛋白rROP38的反应原性进行鉴定。方法根据已发表的ROP38基因序列设计引物，通过RT?PCR方法扩增弓形虫RH株的ROP38基因。将ROP38部分基因克隆到原核表达载体pET?28a（＋）后，重组质粒转化大肠杆菌BL21（DE3）感受态细胞，IPTG诱导表达并优化表达条件，SDS?PAGE分析表达情况，并通过Western blot鉴定其反应原性。结果SDS?PAGE显示在上清和包涵体均有rROP38表达，分子量约为43kD。Western blot结果显示，rROP38蛋白能被His标签抗体和感染弓形虫的人阳性血清识别，说明ROP38具有良好的反应原性，可以作为潜在的血清学诊断抗原。结论本研究成功获得了具有良好反应原性的弓形虫重组蛋白rROP38。

Prokaryotic expression and identification of rhoptry protein 38 of Toxoplasma gondii

Objective To explore the biological function of rhoptry protein 38（ROP38）of Toxoplasma gondii，and to iden?tify the reactogenicity of the recombinant protein（rROP38）. Methods The ROP38 was amplified by RT?PCR from T. gondiiRH strain，and was cloned into prokaryotic expression vector pET?28a（+）. The recombinant plasmid was transformed into E. co?li BL21（DE3）competent cells. Then the rROP38 was analyzed by SDS?PAGE and identified by Western blot. Results SDS?PAGE showed that rROP38 was efficient expression with a molecular weight of about 43 kD. Western blot showed that rROP38reacted with antibody of His tag or human positive antibody，which indicated that ROP38 had good reactogenicity and could be aserological diagnostic antigen. Conclusion The study successfully obtains the rROP38 of T. gondii with good reactogenicity.