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弓形虫RH株微线体蛋白MIC3的原核表达及其免疫反应性

, PP. 786-789

Keywords: 弓形虫,MIC3,基因表达,免疫反应性

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Abstract:

目的克隆表达弓形虫RH株MIC3基因,并对重组蛋白进行免疫反应性分析。方法应用PCR技术扩增弓形虫RH株MIC3基因,将目的基因片段分别克隆至pET28a(+)和pET32a(+)两种表达载体中构建重组质粒,重组质粒转化至大肠杆菌体BL21(DE3)中,经IPTG诱导表达后,表达产物进行SDS-PAGE电泳分析和Western-blot鉴定。结果成功从弓形虫RH株基因组DNA中克隆出大小约1080bpMIC3基因片段;重组质粒pET28a-MIC3和pET32a-MIC3经过酶切和PCR鉴定,及测序分析,表明重组质粒构建正确。两种重组质粒分别诱导表达后进行SDS-PAGE电泳分析,诱导表达产物分别在50KD及70KD左右出现目的条带,Western-blot分析显示重组蛋白对弓形虫慢性感染小鼠血清具有特异免疫反应性。结论原核表达了弓形虫MIC3重组蛋白,所表达的重组蛋白具有免疫反应性,为下一步利用重组蛋白进行弓形虫病的诊断和疫苗研究奠定基础。

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