GST-AEP融合蛋白的表达、纯化和鉴定  

Expression,Purification and Identification of Glutathione-Anti-epilepsy Peptide (GST-AEP) Fusion Protein

作　　者：高碧峰[1] 王宗仁[1] 卓鹏展 张德安[1] 肖茜[1] 

机构地区：[1]第四军医大学附属西京医院中医科,西安710032 [2]西安铁路医院内一科,西安710004

出　　处：《中国生物制品学杂志》2007年第7期497-499,共3页Chinese Journal of Biologicals

基　　金：国家自然科学基金资助项目(30671764)

摘　　要：目的利用GST融合基因表达系统表达GST-AEP融合蛋白,并进行纯化及鉴定。方法IPTG诱导重组质粒pGEX-4T-1/AEP在大肠杆菌BL21(DE3)中表达,溶菌酶裂解后,采用GST蛋白纯化系统进行纯化,所得产物进行12%SDS-PAGE及Western blot鉴定。结果大肠杆菌经诱导,高效表达出相对分子质量约34000的蛋白,与GST-AEP融合蛋白相符。Western blot结果显示该蛋白能够被兔抗GST多克隆抗体识别。结论已成功表达并纯化了融合蛋白,为更深入研究AEP的结构和功能奠定了基础。Objective To express,purify and identify GST-AEP fusion protein. Methods Induce the expression of recombinant plasmid pGEX-4T-1/AEP in E. coil BL21 （ DE3 ） by IPTG, lyze the bacteria with lysozyme and purify the expressed product by GST protein purification system. Identify the expressed fusion protein by SDS-PAGE and Western blot. Results The relative molecular mass of expressed product was 34 000, which was consistent with that of GST-AEP fusion protein. Western blot proved that the expressed fusion protein was recognized by rabbit anti-GST polyclonal antibody. Conclusion GST-AEP fusion protein was successfully expressed and purified, which laid a foundation of further study on structure and function of AEP.

关 键 词：抗癫痫肽 谷胱甘肽 融合蛋白 表达 纯化 

分 类 号：Q786[生物学—分子生物学]