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.
出处
《吉林大学学报（理学版）》
CAS
CSCD
北大核心
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)
关键词
糖尿病
血栓
促胰岛素水蛭素
表达
纯化
功能分析
diabetes mellitus
thrombosis
5rolGLP-HV
expression
purification
function analysis
