Expression, purification and bioactivity determination of epidermal growth factor receptor protein tyro-sine kinase
文献类型：期刊论文
| 作者 | Li Lin1; Zhao Haidan1; Li Guowei2; Chen Jing2 ; Shen Xu2; Mei Changlin1
|
| 刊名 | Academic Journal of Second Military Medical University
 |
| 出版日期 | 2004 |
| 卷号 | 25期号:12页码:1353-1356 |
| 关键词 | epidermal growth factor receptor tyrosine kinase gene expression |
| ISSN号 | 0258-879X |
| 其他题名 | 表皮生长因子受体蛋白酪氨酸激酶的表达、纯化及活性测定 |
| 文献子类 | Article |
| 英文摘要 | Objectivc: To express the epidermal growth factor receptor (EGFR) protein tyrosine kinase(RTK) in E. colt and to detect its bioactivity after purifying the fusion protein. Methods: Using the plasmid pRK5 containing the EGFR cDNA as the template, we selectively amplified the fragment which coding EGFR-RTK. The fragment was then inserted into the prokaryotic expression vector (pQE30) and transformed into competent E. coli cells (M15). Fusion protein expression was induced by IPTG. After renaturation, the protein was purified by affinity chromatography and the bioactivity was examined by ELISA. Results: We successfully inserted EGFR-RTK cDNA fragment of 933 bp into vector pQE30 and constructed the expression vector pQE30-RTK. The fusion protein, with a molecular weight of 37 000, was efficiently expressed as inclusion in the prokaryotic expressive system with the yield of 65. 2% total bacterial protein. After renaturation and purification, the recombinant protein was proven to be bioactive by ELISA. Conclusion: The expression and purification of the bioactive EGFR-RTK in prokaryotic expression system in this study is cheaper and easier than the traditional insect expression system. |
| 资助项目 | 上海市科委重大基础项目基金[00000000] ; 国家自然科学基金[00000000] ; 国家科技部重大科技专项基金[00000000] |
| WOS研究方向 | Dentistry, Oral Surgery & Medicine (provided by Clarivate Analytics) |
| 语种 | 中文 |
| CSCD记录号 | CSCD:1805185 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/268415]  |
| 专题 | 药物安全性评价中心 |
| 作者单位 | 1.Department of Nephrology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.; 2.Shanghai Institute of Materia Meclica, Chinese Academy of Sciences, Shanghai 200023, China. |
| 推荐引用方式 GB/T 7714 | Li Lin,Zhao Haidan,Li Guowei,et al. Expression, purification and bioactivity determination of epidermal growth factor receptor protein tyro-sine kinase[J]. Academic Journal of Second Military Medical University,2004,25(12):1353-1356. |
| APA | Li Lin,Zhao Haidan,Li Guowei,Chen Jing,Shen Xu,&Mei Changlin.(2004).Expression, purification and bioactivity determination of epidermal growth factor receptor protein tyro-sine kinase.Academic Journal of Second Military Medical University,25(12),1353-1356. |
| MLA | Li Lin,et al."Expression, purification and bioactivity determination of epidermal growth factor receptor protein tyro-sine kinase".Academic Journal of Second Military Medical University 25.12(2004):1353-1356. |
入库方式： OAI收割
来源：上海药物研究所
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