Construction of Cardiac-specific Eukaryotic Expression Vector and Its Identification in vitro
YU Qin ZHENG Xiaoqun,FANG Weiyi.Dalian Medical University,Dalian 116023;Department of Cardiology,Shanghai Chest Hospital,Shanghai 200030
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 cardiomyocytes. Conclusion: the eukaryotic expression vector with MLC-2v promoter is a cardiac-specific expression vector.
【CateGory Index】： Q78