双功能促胰岛素水蛭素(5rolGLP-HV)的构建及功能分析  

Construction and Function Analysis of Dual-functional Peptide(5rolGLP-HV)

作　　者：张耀方[1,2] 马百成[1] 屠培培[1] 李宠[1] 马志华[1] 姬艳丽[1] 董震[1] 马超[1] 郝军凤 李晓丹[1] 王宇[1] 李明刚[1] 

机构地区：[1]南开大学生命科学学院分子生物学研究所生物活性材料教育部重点实验室,天津300071 [2]天津农学院,天津300384

出　　处：《吉林大学学报（理学版）》2014年第1期131-136,共6页Journal of Jilin University:Science Edition

基　　金：国家高技术研究发展计划863项目基金(批准号:2008AA02Z205)

摘　　要：采用分子克隆方法获得5拷贝口服长效促胰岛素多肽(rolGLP-1)和水蛭素(rHV)融合基因促胰岛素水蛭素(5rolGLP-HV),并将该基因克隆至pET22b(+)表达载体上,转化大肠杆菌BL21(DE3),实现了高效表达.表达产物经SDS-PAGE电泳及Western blot鉴定后用Ni-NTA亲和层析柱纯化,得到高纯度的5rolGLP-HV.用STZ诱导Ⅱ型糖尿病小鼠和血栓模型鼠进行药物功能分析,结果表明,5rolGLP-HV有较好的降糖和抗血栓效果.We constructed a fusion gene consisted of five copies long-acting Glucagon-like peptide-1 （rolGLP-1）and Hirudin （rHV）called 5rolGLP-HV by molecular cloning.5rolGLP-HV gene was cloned into the expression vector pET22b （＋）and translated into BL21 （DE3 ）,and a highly expression bacterial strain was obtained.The expression product 5rolGLP-HV was analyzed by SDS-PAGE electrophoresis and Western bolt,and was purified with Ni-NTA.Function analysis was performed via STZ-induced type Ⅱ diabetic mice and thrombosis mice.The results show 5rolGLP-HV has reducing blood glucose and antithrombotic effects.

关 键 词：糖尿病 血栓 促胰岛素水蛭素 表达 纯化 功能分析 

分 类 号：Q189[生物学—神经生物学]