| 人重组载脂蛋白E在大肠杆菌中的表达和纯化 |
| |
| 引用本文： | 国汉邦,董军,王抒. 人重组载脂蛋白E在大肠杆菌中的表达和纯化[J]. 中国动脉硬化杂志, 2002, 10(5): 389-391 |
| |
| 作者姓名： | 国汉邦 董军 王抒 |
| |
| 作者单位： | 卫生部北京医院老年医学研究所,北京市,100730 |
| |
| 基金项目： | 国家重点项目研究发展规划 (G2 0 0 0 0 5 70 0 7)资助 |
| |
| 摘 要： | 用含有载脂蛋白EcDNA的PET32a原核表达载体转化大肠杆菌BL21（DE3），使之高效表达载脂蛋白E-thioredoxin融合蛋白，利用融合蛋白上的一段组氨酸序列，用镍离子亲合层析柱进行分离纯化。由于在载脂蛋白E和thioredoxin之间存在凝血酶的识别位点，用凝血酶消化后，经SephacrylS-300凝胶过滤得到人重组载脂蛋白E。用此方法可以从1L大肠杆菌培养液中纯化20-30mg高纯度的各种载脂蛋白E异构体和非自然存原突变体。方法简便，产量高，纯度达95%以上。
|
| 关 键 词： | 大肠杆菌 载脂蛋白E 重组蛋白 表达 纯化 |
| 文章编号： | 1007-3949(2002)10-05-0389-03 |
| 收稿时间： | 2002-05-27 |
| 修稿时间： | 2002-05-27 |
Expression and Purification of Human Recombinant Apolipoprotein E from Escherichia coli |
| |
| GUO Han Bang,DONG Jun,and WANG Shu. Expression and Purification of Human Recombinant Apolipoprotein E from Escherichia coli[J]. Chinese Journal of Arteriosclerosis, 2002, 10(5): 389-391 |
| |
| Authors: | GUO Han Bang DONG Jun and WANG Shu |
| |
| Affiliation: | Institute of Geriatrics, Beijing Hospital, Ministry of Health, Beijing 100730, China |
| |
| Abstract: | Aim To separate and purify human recombinant apolipoprotein E (Apo E) from Escherichia coli strain BL21(DE3). Methods PET32a Apo E constructs were transformed into BL21 and protein expression was induced. Apo E thioredoxin fusion protein was separated by Ni 2+ affinity column. After digestion of thioredoxin by thrombin, Apo E was purified by Sephacryl S 300 gel filtration column. Results Apo E2, Apo E3 and Apo E4 were produced by this method, with high yield and high purity. Conclusions A new system for separation and purification of human recombinant Apo E was successfully established. |
| |
| Keywords: | Apolipoprotein E Recombinant Protein Expression Purification |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
| 点击此处可从《中国动脉硬化杂志》浏览原始摘要信息 |
|
点击此处可从《中国动脉硬化杂志》下载全文 |
