| 半乳糖凝集素-1的表达纯化与生物活性分析 |
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| 引用本文： | 易绍琼,任声权,于婷,张晓艳,刘树玲,陈薇.半乳糖凝集素-1的表达纯化与生物活性分析[J].军事医学科学院院刊,2009,33(3):231-233. |
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| 作者姓名： | 易绍琼 任声权 于婷 张晓艳 刘树玲 陈薇 |
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| 作者单位： | 军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京,100071 |
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| 摘 要： | 目的：获得重组人半乳糖凝集素-1（galectin-1）蛋白,并分析其生物学活性。方法：PCR方法扩增目的基因,将其克隆到原核表达载体pET21a（＋）,获得重组质粒pET-galectin;将其转化至表达宿主菌BL21（DE3）中,用IPTG进行诱导,获得目的蛋白的表达;采用Ni-NTA亲和层析柱对目的蛋白进行纯化,用红细胞凝集试验分析其生物学活性。结果与结论：成功构建重组原核表达质粒pET-galectin,目的蛋白在大肠杆菌中获得高效表达,经Ni-NTA亲和层析柱一步纯化后,得到的蛋白纯度可达80%以上。红细胞凝集试验结果表明获得的重组蛋白具有较好的生物学活性,可进一步用于半乳糖凝集素-1功能研究。
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| 关 键 词： | 半乳糖凝集素 原核表达 蛋白纯化 红细胞凝集试验 |
Expression, purification and bioactivity analysis of galectin-1 |
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| YI Shao-Qiong,REN Sheng-Quan,YU Ting,ZHANG Xiao-Yan,LIU Shu-Ling,CHEN Wei.Expression, purification and bioactivity analysis of galectin-1[J].Bulletin of the Academy of Military Medical Sciences,2009,33(3):231-233. |
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| Authors: | YI Shao-Qiong REN Sheng-Quan YU Ting ZHANG Xiao-Yan LIU Shu-Ling CHEN Wei |
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| Institution: | ( State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China) |
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| Abstract: | Objective：To obtain recombinant human galectin-1 and to investigate its bioactivity. Methods： The target gene galectin-1 was amplified by PCR and then cloned into the plasmid pET21a （ ＋ ）. The recombinant plasmid pET-galectin was transformed into host BL21 （ DE3 ）. Galectin-1 was expressed well in BL21 after induction by IPTG. With a 6 × His tag at its C terminal, the galectin-1 was purified by Ni-NTA chromatography. The lectin activity of galectin-1 was determined by a hemagglutination assay. Results and Conclusion：The recombinant protein was expressed at a high level in BL21. A highly purified target protein was obtained by Ni-NTA chromatography, which could agglutinate mouse erythrocytes at 30 μg/ml. |
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| Keywords: | galectin prokaryotic expression purification hemagglutination assay |
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