抗癫痫肽/GST融合表达及包涵体蛋白复性纯化  

Expression and Purification of the Fusion Protein Anti-epilepsy Peptide/Glutathiones-transferase

作　　者：张德安[1] 王宗仁[1] 邵中军[2] 高碧峰[1] 李晶华[1] 马静[1] 

机构地区：[1]第四军医大学附属西京医院,西安710032 [2]第四军医大学

出　　处：《浙江中医药大学学报》2007年第1期38-40,共3页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金资助项目(No:30671764)~~

摘　　要：[目的]获得高纯度的谷胱甘肽硫转移酶/抗癫痫肽(GST-AEP)融合蛋白。[方法]异丙基-β-D硫代半乳糖苷(IPTG)诱导重组大肠杆菌DH5α表达GST-AEP融合蛋白,融合蛋白包涵体经过洗涤、变性、复性后,通过12%SDS-PAGE凝胶电泳,考马斯亮蓝染色,凝胶薄层扫描检测其纯度,透析管进一步纯化蛋白,BCA法定量。[结果]从融合蛋白包涵体中获得了纯度达90%以上的目的蛋白,定量约0.163μg/μl。[结论]建立了有效纯化融合蛋白包涵体GST-AEP的方法,为对AEP进一步的相关研究奠定了基础。[Aim] To gain the fusion protein purified GST-AEP. [Methods] The protein GST-AEP was expressed in E. Coli-DH5α as a fusion protein induced by IPTG. The protein was a kind of inclusion body. The purifying and refolding to inclusion body were optimized. The purity of GST-AEP was identified by 12% SDS-PAGE and thin-layer scanning analysis. The quantitation of the fusion protein GST-AEP was done with BCA Protein Assay. [Results] Purity of GST-AEP was higher than 90% and concentration was about 0. 163μg/μl. [Conclusion] The fusion protein was highly purified and the method of fusion protein purification from the inclusion body was developed,which was the basis for further study on AEP.

关 键 词：抗癫痫肽 融合蛋白 包涵体 复性 蛋白纯化 

分 类 号：R392.9[医药卫生—免疫学]