| 羟胺切割Neurturin融合蛋白表达载体的构建、表达产物纯化及生物活性 |
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| 引用本文： | 李鸿钧,马雁冰,戴长柏,孙茂盛.羟胺切割Neurturin融合蛋白表达载体的构建、表达产物纯化及生物活性[J].中国生物化学与分子生物学报,2001,17(2):139-142. |
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| 作者姓名： | 李鸿钧 马雁冰 戴长柏 孙茂盛 |
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| 作者单位： | 中国医学科学院中国协和医科大学医学生物学研究所 |
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| 摘 要： | Neurturin (NTN)是新近发现的一种神经营养因子 ,是 GDNF家族的成员之一 .将 5′端引入了羟胺切割位点的 h NTN基因克隆到硫氧还蛋白融合表达载体 p Thio His A,在宿主菌 BL2 1中获得了稳定、高效表达 ,表达产物以包涵体形式存在 .在变性条件下经羟胺切割、柱层析纯化后复性 ,获得纯度达 90 %以上的 rh NTN.经鸡胚背根神经节 (DRG)培养法测定具有生物学活性 .
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| 关 键 词： | neurturin 融合蛋白 羟胺切割 鸡胚背根神经节 纯化 生物活性 |
| 收稿时间： | 2001-04-20 |
| 修稿时间： | 2000年9月11日 |
Construction of Neurturin Fusion Expression Vector with Hydroxylamine Site and Purification and Bioactivity of Expressive Products |
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| LI Hong jun,MA Yan bing,DAI Chang bai,SUN Mao sheng.Construction of Neurturin Fusion Expression Vector with Hydroxylamine Site and Purification and Bioactivity of Expressive Products[J].Chinese Journal of Biochemistry and Molecular Biology,2001,17(2):139-142. |
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| Authors: | LI Hong jun MA Yan bing DAI Chang bai SUN Mao sheng |
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| Institution: | (Institute of Medical Biology,CAMS and PUMC,Kunming 650118,China |
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| Abstract: | Neurturin (NTN) is a novel neurotrophic factor,which is one of the GDNF family members.The 5′ modified hNTN gene was cloned to plasmid pThioHisA to construct the thioredoxin (Trx) and hNTN fusion expression vector with hydroxylamine site between the two fused genes.Higher levels of expression of hNTN fusion protein was obtained in E.coli BL21.The expressed fusion protein existed in inclusion body.About 90% pure rhNTN was obtained after cleavaging with hydroxylamine in denatured condition and purifying with SP Sepharose Fast Flow column and renaturing.The bioactivity was tested by dorsal root ganglia (DRG) of chicken embryo culture. |
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| Keywords: | neurturin fusion protein hydroxylamine cleavage dorsal root ganglia purification bioactivity |
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