| 蛇神经毒素在毕赤酵母中的分泌表达及其分离纯化 |
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| 引用本文： | 张伟,陈劲春.蛇神经毒素在毕赤酵母中的分泌表达及其分离纯化[J].北京化工大学学报(自然科学版),2005,32(6):20-22. |
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| 作者姓名： | 张伟 陈劲春 |
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| 作者单位： | 北京化工大学生命科学与技术学院，北京 100029 |
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| 摘 要： | 文中主要研究了毕赤酵母基因工程菌的构建及其发酵表达产物蛇神经毒素的分离纯化条件。蛇神经毒素编码基因由本实验室设计、合成，并重组构建成表达质粒pPIC9k-hPK。经电穿孔将该质粒转化毕赤酵母GS115 (His-Mut+)，筛选His+Muts型菌株，经诱导表达，产物进行SDS- PAGE电泳鉴定，相对分子质量与理论值一致，目标蛋白经小试发酵产量可达350mg/L。利用超滤、离子交换树脂及分子筛分离纯化，产物经高效液相色谱(HPLC)分析纯度达92%。
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| 关 键 词： | 毕赤酵母 分离纯化 神经毒素 毕赤酵母 分离纯化 神经毒素 |
| 收稿时间： | 2004-12-31 |
| 修稿时间： | 2004年12月31日 |
Separation and purification of recombinant snake neurotoxins from Pichia pastoris |
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| ZHANG Wei,CHEN Jin-chun.Separation and purification of recombinant snake neurotoxins from Pichia pastoris[J].Journal of Beijing University of Chemical Technology,2005,32(6):20-22. |
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| Authors: | ZHANG Wei CHEN Jin-chun |
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| Institution: | College of Life Science and Technology,Beijing University of Chemical Te chnology, Beijing 100029, China |
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| Abstract: | The conditions of fermentation for recombinant neurotoxins tranfering into P Pastoris GS 115 and their further purification procedures were investigated. An identification of the target protein was carried out with the SDS-PAGE electrophoresis and its productivity reached 350 mg/L. After the purification procedures in- cluding the ultra filtration, ion-exchange chromatography and the gel filtration were performed, the purity of the product, which was analyzed by HPLC, was 92 %. |
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| Keywords: | Pichia pastoris separation and purification neurotoxins |
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