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。
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 BI21 （DE3）. GST/AEP fusion protein was expressed under IFTG 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 B121（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 BI21 （DE3）was expressed.
出处
《现代生物医学进展》
CAS
2006年第8期1-3,共3页
Progress in Modern Biomedicine
基金
国家自然科学基金资助项目(30671764)
关键词
抗癫痫肽
GST-融合蛋白
原核表达
基因扩增
Anti - epilepsy peptide（AEP）
GST/AEP fusion protein
Prokaryotic expression
Gene amplification
