心肌特异性hVEGF_(165)真核表达载体的构建及鉴定  

Construction of Cardiac-specific Eukaryotic Expression Vector and Its Identification in vitro

作　　者：于勤[1] 郑晓群[1] 方唯一[2] 

机构地区：[1]大连医科大学附属中山医院,辽宁大连116023 [2]上海市胸科医院心内科,上海200030

出　　处：《中国临床医学》2006年第2期167-169,共3页Chinese Journal of Clinical Medicine

摘　　要：目的:构建MLC-2v启动子驱动的真核表达载体并鉴定特异性。方法:用MLC-2v启动子基因片段替代质粒 pIRES2-EGFP的CMV启动子,而保留其增强子序列,并将目的基因hVEGF165分别克隆到两种启动子驱动的质粒,构建 MLC-2v／pIRES2-EGFP-hVEGF165和pIRES2-EGFP-hVEGF1652种质粒,瞬时转染心肌、内皮、平滑肌和成纤维细胞。结果: 转染pIRES2-EGFP-hVEGF165,4种细胞均表达、目的基因及产物;而转染MLC-2v／pIRES2-EGFP-hVEGF165,仅心肌细胞表达GFP和目的基因产物。结论:MLC-2v启动子驱动的质粒可心肌特异表达目的基因及蛋白。Objective：To construct and identify a cardiac-specific eukaryotic expression vector Methods： To construct two eukaryotic expression vectors. The hVEGF165 DNA fragment was inserted into a enhanced green fluorescent protein fusion vector with CMV promoter and the other CMV promoter was replaced with MLC-2v promoter preserving CMV enhancer, the latter is cardiac-specific promoter. The two different vectors were transfected into endothelial cells , cardiomyocytes, smooth cells and fibroblast cells with liposome. Results： The four cultivated cells were successfully transfected with pIRES2-EGFP- hVEGF165, which confirmed by fluoroscopy, biochemistry and immunohistochemistry, but the vector with MLC-2v promoter only express the target gene in eardiomyocytes, Conclusion： the eukaryotie expression vector with MLC-2v promoter is a cardiac-specific expression vector.

关 键 词：质粒 肌凝蛋白轻链-2v 启动子 心肌细胞 特异性表达 

分 类 号：Q782[生物学—分子生物学]