重组人胰高血糖素样肽-1类似物的制备及活性分析
Preparation and preliminary analysis of activity of recombinant GLP-1 analogue
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摘要: 通过重叠PCR的方法获得重组人胰高血糖素样肽-1(GLP-1)类似物PP-hGly8GLP-1的目的基因片段，将该基因与融合表达载体pED连接，并在融合伴侣的N末端插入6×His标签，转入E.coli BL21(DE3)，构建出重组GLP-1类似物融合表达菌。PP-hGly8GLP-1通过Ni离子亲和色谱和酸水解等步骤分离纯化得到，其纯度大于90%，用电喷雾质谱法测定其相对分子质量与理论分析结果一致。ICR小鼠经口给予耐血糖及血浆胰岛素测定的结果表明，PP-hGly8GLP-1具有明显的降血糖活性和促胰岛分泌作用（P<0.01），为开发潜在的治疗糖尿病药物提供了理论基础。Abstract: The PP-hGly8GLP-1 gene was obtained by overlap PCR and cloned into fusion expression vector pED containing AnsB-C gene and 6×His-tag gene at the N-terminus of AnsB-C gene.The recombinant plasmid was transformed into E.coli BL21(DE3).Recombinant GLP-1 analogue peptide PP-hGly8GLP-1 was purified by Nickle ion affinity chromatography and hydrochloric acid hydrolysis.The purity of recombinant PP-hGly8GLP-1 product is more than 90%.Its molecular weight (MW) was determined by electrospray ionization mass spectrometry,with the result similar to that of the theoretical molecular weight.The results of oral glucose tolerance test and plasma insulin measurement with ICR mice showed that the recombinant GLP-1 analogue peptide has significant effects in decreasing glycemic activity and promoting insulin secretion(P<0.01).It would be a powerful potential medicine for the treatment of type 2 diabetes.
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