| 重组人bFGF蛋白的克隆、表达、纯化及鉴定 |
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| 引用本文： | 王辉,刘岱琳,张英起.重组人bFGF蛋白的克隆、表达、纯化及鉴定[J].武警医学院学报,2010,19(2):88-89,92,F0004. |
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| 作者姓名： | 王辉 刘岱琳 张英起 |
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| 作者单位： | 1. 武警医学院基础医学实验室,天津,300162
2. 第四军医大学药学系生物技术中心,陕西,西安,710032 |
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| 摘 要： | 【目的】构建人碱性成纤维细胞生长因子（bFGF）基因的原核表达载体,并对其进行诱导表达和蛋白质纯化,以获得大量重组人bFGF蛋白。【方法】人工合成bFGF基因,进行测序分析,将该基因克隆入原核表达载体pET22b中,转化感受态细胞大肠杆菌RPX,经IPTG诱导表达重组bFGF蛋白,对表达产物进行SDS-PAGE电泳分析和Western blotting检测分析。【结果】成功构建了人重组bFGF表达质粒pET22b-rhbFGF,表达的蛋白经SDS-PAGE电泳分析,分子量约在18×103Da处出现了一条新生的蛋白条带,经灰度扫描检测,表达量约占菌体总蛋白的28%,纯化后的蛋白质灰度扫描检测,纯度为95%。【结论】成功构建了重组人bFGF原核表达载体,并成功纯化了该基因的原核表达产物,为下一步人bFGF的产业化打下了基础。
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| 关 键 词： | bFGF 成纤维细胞 基因重组 表达 纯化 载体 |
Identification,purification,expression and cloning of fusion protein recombinant human bFGF |
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| WANG Hui,LIU Dai-lin,ZHANG Ying-qi.Identification,purification,expression and cloning of fusion protein recombinant human bFGF[J].Acta Academiae Medicinae CPAPF,2010,19(2):88-89,92,F0004. |
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| Authors: | WANG Hui LIU Dai-lin ZHANG Ying-qi |
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| Institution: | WANG Hui,LIU Dai-lin,ZHANG Ying-qi(Medical College of Chinese People\'s Armed Police Force,Tianjin 300162,China) |
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| Abstract: | Objective]To construct the recombinant expression vector of human bFGF(basic fibroblast growth factor)gene and to purify and identify the protein rhbFGF.Methods]A 465 bp of human bFGF gene fragment was synthesized and cloned into Pet22b vector,an E.coli expression vector.The plasmid was transformed into E.coli RPX and induced to express protein rhbFGF with IPTG.The expression of bFGF was detected by SDS-PAGE and Western blotting.Results]A novel protein with expected molecular mass was expressed upon indu... |
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| Keywords: | bFGF |
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