细胞与分子免疫学杂志 , Chinese Journal of Cellular and Molecular Immunology, 编辑部邮箱 , 2011年10期 【作者】 尹志奎; 邓智建; 郑斌;
【Author】 YIN Zhi-kui1, DENG Zhi-jian1, ZHENG Bin2* 1College of Pharmacy, 2Department of Parasitology, Xinxiang Medical University, Xinxiang 453003, China
【机构】 新乡医学院药学院办公室; 新乡医学院寄生虫学教研室;
【摘要】 目的:体外制备弓形虫醛缩酶(aldolase)蛋白及其多克隆抗体。方法:以弓形虫cDNA链为模板,PCR扩增醛缩酶基因,构建aldolase/pET30a原核表达系统;IPTG诱导表达aldolase-His6蛋白;用纯化的蛋白加免疫佐剂免疫SD大鼠,收集抗血清,制备多克隆抗体,亲和层析纯化并分析抗体的效价。结果:构建了aldolase原核表达系统,表达并纯化了aldolase-His6蛋白;获得了抗该蛋白的大鼠源性抗血清,纯化后的多克隆抗体效价为1∶4000。结论:在体外制备并纯化了aldolase-His6蛋白及其多克隆抗体,为后续研究aldolase的功能奠定了基础。 更多还原
【Abstract】 AIM: To produce the aldolase protein and its polyclonal antibody. METHODS: Aldolase gene was obtained from cDNA library by PCR amplification and subcloned to vector pET30a. The recombinant protein aldolase-His6 was expressed in E.coli upon IPTG induction and then purified with affinity chromatography. The purified protein mixed with adjuvant was used to immunize SD rats to produce the polyclonal antibodies. RESULTS: The recombinant plasmid aldolase/pET30a was constructed successfully and express... 更多还原 【关键词】 刚地弓形虫; 醛缩酶; 蛋白表达; 多克隆抗体; 【Key words】 Toxoplasma gondii; aldolase; protein expression; polyclonal antibody; 【基金】 河南省科技攻关计划项目(112102310209);新乡医学院博士科研启动基金资助(2009)【分类号】R392.1
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