| pPRMETb-TRAIL原核表达载体的构建和表达及纯化 |
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| 引用本文： | 孙等军,方琴,陈剑,肖芸,任志娟,张维,张旭,王季石. pPRMETb-TRAIL原核表达载体的构建和表达及纯化[J]. 贵阳医学院学报, 2002, 27(1): 4-6,10 |
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| 作者姓名： | 孙等军 方琴 陈剑 肖芸 任志娟 张维 张旭 王季石 |
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| 作者单位： | 1. 贵阳医学院附院药剂科,贵州贵阳,550004
2. 贵阳医学院附院血液内科,贵州贵阳,550004 |
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| 基金项目： | 贵州省省长专项资金资助项目(S2 0 0 1 - 1 4 ) |
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| 摘 要： | 目的：为获取较高纯度的肿瘤坏死因子相关凋亡诱导配体（TNF－related apoptosis-inducing ligand,TRAIL）工程蛋白。方法：将TRAIL基因构建于原核表达载体pPRMETb中，在大肠杆菌中由异丙基硫代β-D-半乳糖苷（IPTG）诱导表达，对包涵体进行洗涤、尿素裂解、复性后过镍柱层析纯化。结果：经酶切、测序鉴定证实pPRMETb-TRAIL构建安全正确，在大肠杆菌xl1-blue中经IPTG诱导表达量较高，包涵体经尿素裂解后行镍柱纯化得到了较高纯度的TRAIL蛋白。结论：成功地建立了获取高纯度TRAIL工程蛋白的技术路线。
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| 关 键 词： | 大肠杆菌 细胞包涵体 聚合酶链反应 肿瘤坏死因子相关凋亡诱导配体 pPRMETb－TRAIL 原核表达载体 |
| 文章编号： | 1000-2707(2002)01-0004-03 |
The Construction, Expression and Purification of Prokaryotic Expression Vector pPRMETb-TRAIL |
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| SUN Deng jun ,FANG Qin ,CHEN Jian ,XIAO Yun ,REN Zhi juan ,ZHANG Wei ,ZHANG Xu ,WANG Ji shi. The Construction, Expression and Purification of Prokaryotic Expression Vector pPRMETb-TRAIL[J]. Journal of Guiyang Medical College, 2002, 27(1): 4-6,10 |
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| Authors: | SUN Deng jun FANG Qin CHEN Jian XIAO Yun REN Zhi juan ZHANG Wei ZHANG Xu WANG Ji shi |
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| Affiliation: | SUN Deng jun 1,FANG Qin 1,CHEN Jian 2,XIAO Yun 2,REN Zhi juan 2,ZHANG Wei 2,ZHANG Xu 2,WANG Ji shi 2 |
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| Abstract: | Objective: To abtain a pure TRAIL protein. Methods: A prokaryotic expression vector pPRMETb TRAIL was constructed and induced by IPTG to express in E.coli. TRAIL protein was purified with nickel column chromatography after inclusion body lysis , protein renaturation and recovery . Results:The constrution of prokaryotic expression vector pPRMETb TRAIL was proofed correct by digestive identification and sequence measuring. TRAIL protein induced by IPTG had a high expression in xl1 blue. A pure TRAIL protein was obtained with nickel column chromatography after inclusion body lysis. Conclusion: A technological procedure of obtaining a pure TRAIL protein was successfully established. |
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| Keywords: | escherichia coli cellular inclusions polymexase chain reaction TNF related poptosis inducing ligand |
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