| 重组大鼠细胞球蛋白的制备、纯化与活性 |
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| 引用本文： | 张亭亭,李招发,许瑞安. 重组大鼠细胞球蛋白的制备、纯化与活性[J]. 华侨大学学报(自然科学版), 2009, 30(6) |
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| 作者姓名： | 张亭亭 李招发 许瑞安 |
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| 作者单位： | 华侨大学,分子药物学研究所,福建,泉州,362021;华侨大学,分子药物学研究所,福建,泉州,362021;华侨大学,分子药物学研究所,福建,泉州,362021 |
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| 基金项目： | 国家高新技术研究发展(863)计划项目(2005216060,2008AA022135);;福建省自然科学基金资助项目(2007J0105);;福建省科技厅重点项目(2008Y0084) |
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| 摘 要： | 通过聚合酶链式反应(PCR)克隆鼠源细胞球蛋白(Cytoglobin,CYGB)基因,构建原核表达载体pET22b-Cygb,转入E.coliBL21(DE3)经乳糖诱导表达CYGB,发现CYGB主要以可溶形式存在,其表达量占可溶性总蛋白的35%以上.通过DEAE阴离子交换柱层析和Sephacryl S-100凝胶过滤层析,CYGB纯度可超过95%.重组表达的可溶性CYGB不但具有过氧化物酶活性,其活性为(3.23±0.12)mkat.(g.min)-1,还能保护胎鼠原代皮肤成纤维细胞减轻氧化应激损伤(H2O2模型).
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| 关 键 词： | 重组细胞球蛋白 高效可溶表达 分离纯化 鉴定 活性检测 |
Preparation, Purification and Activity of Recombinant Rat Cytoglobin |
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| ZHANG Ting-ting,LI Zhao-fa,XU Rui-an. Preparation, Purification and Activity of Recombinant Rat Cytoglobin[J]. Journal of Huaqiao University(Natural Science), 2009, 30(6) |
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| Authors: | ZHANG Ting-ting LI Zhao-fa XU Rui-an |
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| Affiliation: | Institute of Molecular Medicine;Huaqiao University;Quanzhou 362021;China |
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| Abstract: | Rat Cygb cDNA was cloned through PCR with rAAV-Cygb as template and inserted into a prokaryotic expression vector to construct a recombinant plasmid,pET22b-Cygb.Then the recombinant plasmid pET22b-Cygb was transformed into E.coli BL21(DE3) and expressed by lactose induction.The CYGB protein accounted for more than 35% of total soluble protein.The purified CYGB protein was achieved through DEAE anion-exchange chromatography and Sephacryl S-100 gel filtration chromatography,respectively.Eventually,the purity ... |
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| Keywords: | recombinant CYGB soluble high-expression isolation and purification identification activity analysis |
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