| 高碧峰 王宗仁 张德安.GST/ AEP 融合蛋白原核表达载体的构建、表达及鉴定[J].现代生物医学进展英文版,2006,6(8):1-3. |
| GST/ AEP 融合蛋白原核表达载体的构建、表达及鉴定 |
| Construction and expression of prokaryotic expression vector ofGST/ AEP fusion protein and identification |
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| DOI： |
| 中文关键词: 抗癫痫肽 GST- 融合蛋白 原核表达 基因扩增 |
| 英文关键词: Anti- epilepsy peptide( AEP) GST/ AEP fusion protein Prokaryotic expression Gene amplification |
| 基金项目:国家自然科学基金资助项目( 30671764) |
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| 中文摘要: |
| 目的: 为进一步研究抗癫痫肽( Anti- epilepsy peptide, AEP) 的抗痫机制及筛选其相关作用蛋白, 进行GST/ AEP 融合蛋
白原核表达载体的构建及融合蛋白的表达。方法: 通过PCR 基因扩增对AEP 基因进行扩增, 并将其克隆于谷胱甘肽- S- 转移
酶( GST) 融合蛋白表达质粒pGEX- 4T- 1 中, 经酶切、序列鉴定分析后, 用该重组质粒转化大肠杆菌Bl21( DE3) , 经IPTG 诱导获得
表达, 并采用Western Blot 进行检测。结果: 成功构建了AEP 原核表达载体, 并在大肠杆菌Bl21 中获得表达。结论: 成功构建了
GST/ AEP 原核表达载体, 并表达了GST/ AEP 融合蛋白。 |
| 英文摘要: |
| Objective: To express fusion protein of GST and Anti- epilepsy peptide ( AEP) in E. coli. Methods: The core fragment of
AEP was cloned into pGEX- 4T- 1 cotaining glutathione s- transterase( GST) fusion protein gene. Following the restriction enzyme digestion
analysis and sequencing, pGEX- 4T- 1/ AEP was transformed into E. coli Bl21( DE3) . GST/ AEP fusion protein was expressed under IPTG induction
and the AEP protein was identified by the Western- blot. Results: The restriction endonuclease digestion analysis of recombinant plasmid
demonstrated that the AEP gene had been exactly inserted in pGEX- 4T- 1, SDS- PAGE analysis showed that the relative molecular mass of the
fusion protein was about 34KD. The GST/ AEP fusion protein was expressed in E. coli Bl21( DE3) and identified by the Western- blot. Conclusion:
pGEX- 4T- 1/ AEP vector was successfully constructed, and the GST/ AEP fusion protein in E. coli Bl21( DE3) was expressed. |
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