CAJ | 학술논문

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.
채용분자극륭방법획득5고패구복장효촉이도소다태(rolGLP-1)화수질소(rHV)융합기인촉이도소수질소(5rolGLP-HV),병장해기인극륭지pET22b(+)표체재체상,전화대장간균BL21(DE3),실현료고효표체.표체산물경 SDS-PAGE 전영급 Western blot 감정후용Ni-NTA친화층석주순화,득도고순도적5rolGLP-HV.용 STZ유도Ⅱ형당뇨병소서화혈전모형서진행약물공능분석,결과표명,5rolGLP-HV유교호적강당화항혈전효과.
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.