弓形虫棒状体蛋白ROP38的原核表达与鉴定  

Prokaryotic expression and identification of rhoptry protein 38 of Toxoplasma gondii

作　　者：崔勇[1] 李瑾[1] 王洪法[1] 仲维霞[1] 孙慧[1] 赵桂华[1] 尹昆[1] 徐超[1] 肖婷[1] 张晓玉[1] 于泓 刘学峰 刘功振[1] 

机构地区：[1]山东省医学科学院,山东省寄生虫病防治研究所,济宁272033 [2]山东省临沂市平邑县畜牧局 [3]山东省临沂市畜牧局

出　　处：《中国血吸虫病防治杂志》2016年第5期554-557,共4页Chinese Journal of Schistosomiasis Control

基　　金：国家自然基金青年项目(31502057;81501770)

摘　　要：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p ET-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43k D。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。Objective To explore the biological function of rhoptry protein 38（ROP38）of Toxoplasma gondii,and to identify the reactogenicity of the recombinant protein（rROP38）. Methods The ROP38 was amplified by RT-PCR from T. gondii RH strain,and was cloned into prokaryotic expression vector p ET-28a（＋）. The recombinant plasmid was transformed into E. coli BL21（DE3）competent cells. Then the rROP38 was analyzed by SDS-PAGE and identified by Western blot. Results SDSPAGE showed that rROP38 was efficient expression with a molecular weight of about 43 k D. Western blot showed that rROP38 reacted with antibody of His tag or human positive antibody,which indicated that ROP38 had good reactogenicity and could be a serological diagnostic antigen. Conclusion The study successfully obtains the rROP38 of T. gondii with good reactogenicity.

关 键 词：弓形虫 棒状体蛋白38 原核表达 纳虫空泡 

分 类 号：R531.8[医药卫生—内科学]