【摘 要】
：
目前在弓形虫体内已鉴定出四种微线蛋白复合体(TgMIC1/4/6、TgMIC2/M2AP、TgMIC3/8和TgAMA1/RON2/4/5/8),但新孢子虫微线蛋白的功能及其作用方式未见报道.本研究分别扩增新孢子虫MIC6、MIC1和MIC4基因,构建原核表达质粒pET28a-NcMIC6、pET28a-NcMIC1和pET28a-NcMIC4,表达、纯化重组蛋白rNcMIC6、rNcMIC1和r
【出 处】
：
中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会
论文部分内容阅读
目前在弓形虫体内已鉴定出四种微线蛋白复合体(TgMIC1/4/6、TgMIC2/M2AP、TgMIC3/8和TgAMA1/RON2/4/5/8),但新孢子虫微线蛋白的功能及其作用方式未见报道.本研究分别扩增新孢子虫MIC6、MIC1和MIC4基因,构建原核表达质粒pET28a-NcMIC6、pET28a-NcMIC1和pET28a-NcMIC4,表达、纯化重组蛋白rNcMIC6、rNcMIC1和rNcMIC4.用上述三种重组蛋白分别免疫BALB/c小鼠和新西兰大白兔,制备多克隆抗体,抗体滴度均可达106以上；Western Blotting检测结果显示,anti-rNcMIC6、anti-rNcMIC1和anti-rNcMIC4多抗均分别能与新孢子虫内源性MIC6、MIC1和MIC4发生反应；免疫共沉淀实验结果表明,抗rNcMIC6血清不仅能与新孢子虫内源性MIC6发生反应,同时MIC1和MIC4与其发生共沉淀,说明新孢子虫内源性MIC6与MIC1、MIC4形成蛋白复合体；间接免疫荧光实验显示细胞内新孢子虫的MIC1和MIC6、MIC4和MIC6均共定位于速殖子前端.本研究发现新孢子虫的MIC6、MIC1和MIC4以复合体形式存在,为深入研究新孢子虫MIC蛋白的功能奠定基础.
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