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人αsynuclein突变体的克隆及细菌表面展示
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【免费中医学论文】作者：熊莉丽 赵鹏 郭志云 张建华 茆灿泉【摘要】 目的 在细菌表面构建及展示人αsynuclein突变体A30P、E46K和A53T。方法 采用叠套PCR介导的点突变方法，以野生型αsynuclein为模板。结果 成功克隆了αsynuclein A30P、E46K和A53T 3个突变型基因，并利用细菌亲密素展示系统将野生型αsynuclein及突变体A30P、E46K和A53T成功展示在大肠杆菌表面。结论 αsynuclein及其突变体的细菌表面展示对于αsynuclein空间结构的良好保持和基于αsynuclein神经退行性疾病靶分子药物、分子诊断试剂的研发和αsynuclein蛋白结构与功能的研究等具有重要的意义和价值。
【关键词】 αsynuclein;突变体;生物信息;叠套PCR
【Abstract】 Objective To analyze bioinformatics and display of human αsynuclein mutants of A30P, E46K and A53T proteins on the cell surface of Escherichia coli. Methods Overlap extension PCR, a rapid and efficient method by sitedirected mutagenesis was introduced to clone the mutants A30P, E46K, A53T of αsynuclein. Results All the three cloned mutants, including the wild type of αsynuclein, were successfully displayed on the surface of E. coli strains based on the Enterohemorrhagic E. coli Intimin Eae
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