PbTIP多克隆抗体对恶性疟原虫生长抑制作用的初步研究  

Production of Polyclonal PbTIP Antibody and Its Effect on Growth of Plasmodium falciparum

作　　者：黄豫晓[1] 刘学武[1] 沈燕[1] 李英辉[1] 梁娇[1] 王军[1] 刘忠湘[1] 赵亚[1] 徐志凯[1] 

机构地区：[1]第四军医大学微生物学与病原生物学教研室,西安710032

出　　处：《科学技术与工程》2013年第3期559-563,共5页Science Technology and Engineering

基　　金：国家自然科学基金(81000744);(30901250)资助

摘　　要：首先构建了伯氏疟原虫TIP样蛋白(以下简称PbTIP)的原核表达载体pET32a/PbTIP,在大肠杆菌中成功诱导表达了带His标签的PbTIP融合蛋白。SDS-PAGE分析显示该融合蛋白主要以包涵体形式表达。Western-Blot鉴定虽然证明该融合蛋白表达正确,但即便在变性条件下该蛋白的亲和层析纯化效果也不佳。以PbTIP融合蛋白经过初次和三次加强免疫获得的小鼠PbTIP多克隆抗体滴度达到1∶105以上。进一步的研究表明,小鼠PbTIP抗血清对体外培养恶性疟原虫的生长具有一定的抑制作用,这显示PbTIP多克隆抗体对恶性疟原虫感染具有一定的交叉保护作用。以上实验结果为后续PbTIP在疟原虫免疫抑制中的功能及其机制研究奠定了一定的基础。To preparepolyclonal anti-T cell immunomodulatory protein of Plasmodium berghei and exam effect of it on growth of Plasmodium falciparum.PbTIP gene was constructed in pET32a（＋） vector with a His tag and successfully expressed by E.Coli.The PbTIP-His fusion protein was characterized by SDS-PAGE and Western blots.The polyclonal PbTIP antibody was produced by BALB/c mouse immunized by purified PbTIP-His fusion protein,and the effect of it on growth of Plasmodium falciparum was determined by infection ratio.PbTIP-His fusion protein was successfully expressed in the form of inclusion bodies as evidenced by SDS-PAGE and western blot results.Anti-PbTIP sera were collected in mice enhanced immunized by PbTIP-His protein at a titer of 1：105,which inhibited growth of P.f parasites as evidenced by reduced infection ratio in RBC infected by Plasmodium falciparum.The construction and purification of PbTIP-His fusion protein together with poly-anti-PbTIP could be used for further investigation in Plasmodium berghei,especially for the further study of PbTIP in parasite-host relationship.

关 键 词：疟原虫 T细胞免疫调节蛋白 免疫抑制 

分 类 号：Q591.2[生物学—生物化学]