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.
出处
《浙江中医药大学学报》
CAS
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)~~
