His-Hsp65-6IA2P2的分离纯化及对NOD小鼠降糖作用的初步研究  

Purification of Fusion Protein His-Hsp65-6IA2P2 and Its Preliminary Anti-diabetic Effect Via Nasal Immunization in NOD Mice

作　　者：李国梁[1] 刘坤峰[1] 卢世平 洪流[1] 宗莉[2] 吴洁[1] 曹荣月[1] 金亮[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009 [2]中国药科大学药学院,江苏南京210009

出　　处：《药物生物技术》2015年第4期302-306,共5页Pharmaceutical Biotechnology

基　　金：国家自然科学基金资助项目(No.81172973;31270985);江苏高校优势学科建设工程资助项目(PAPD);mi R-26a靶向TET酶在胰腺成体干细胞分化过程中的作用及机制研究(No.20140029)

摘　　要：构建融合蛋白His-Hsp65-6IA2P2基因工程表达菌E.coli BL21(DE3),诱导其表达可溶性胞外蛋白,并进行初步免疫学研究。构建p ET28a-His-Hsp65-6IA2P2质粒载体,电转化E.coli BL21(DE3),乳糖诱导表达及镍柱亲和层析等纯化步骤后,SDS-PAGE和Band Scan进行检测融合蛋白。最后将此融合蛋白鼻粘膜免疫NOD小鼠,观察其生理特征。融合蛋白His-Hsp65-6IA2P2正确表达,纯化后融合蛋白纯度可达95.2%。免疫动物后能够有效维持正常的体重与血糖。本实验构建了His-Hsp65-6IA2P2基因工程表达菌,成功表达了具有预防治疗及改善1型糖尿病病症潜力的融合蛋白,为进一步探究其药效奠定了基础。To purify soluble extracellular fusion protein His-Hsp65-6IA2P2 expressed by engineering bacteria E. coli BL21（ DE3）and explore its preliminary immunologic study,constructed p ET28a-His-Hsp65-6IA2P2 plasmid vector was electrotransformed into E. coli BL21（ DE3）. After induced by lactose and purified by nickel affinity chromatography etc.,the expression of fusion protein was determined by SDS-PAGE and Band Scan. Finally,nasal immunization in NOD mice was performed. The fusion protein HisHsp65-6IA2P2 was expressed and its purity was up to 95. 2%. Body weight and blood glucose of His-Hsp65-6IA2P2-treated mice were quite normal. Here the expression vector and bacteria for protein His-Hsp65-6IA2P2 was successfully performed. This fusion protein was beneficial for the prevention and treatment of type 1 diabetes symptoms,which laid a foundation for further pharmacological exploration.

关 键 词：融合蛋白 纯化 鼻粘膜免疫 NOD小鼠 1型糖尿病 

分 类 号：Q81[生物学—生物工程]