心肌特异性表达腺病毒载体的构建及鉴定

The construction and identification of cardic specific expression adenovirus vector

摘要: 目的: 构建心肌特异性表达的腺病毒载体,使外源基因在心肌细胞中特异性表达。方法: 将心肌特异性肌凝蛋白轻链蛋白-2(mlc-2v)启动子克隆至腺病毒穿梭质粒,并将绿色荧光蛋白(GFP)基因克隆至此质粒中,同源重组含GFP基因的缺陷型腺病毒(AdmlcGFP),转染心肌细胞及非心肌细胞。结果: 经RT-PCR分析,转染的心肌细胞有GFPmRNA的表达,而转染的非心肌细胞无GFPmRNA的表达;荧光显微镜下观察,可见转染的心肌细胞表达GFP,而转染的的非心肌细胞不表达GFP。结论: 构建的心肌特异性表达腺病毒载体可使外源基因在心肌细胞内特异性表达。

Abstract: Objective: to establish a adenovirus vector that make foreign gene express in cardiomyocytes.Methods: we constructed a recombinant adenovirus （AdmlcGFP） in which contain the GFP gene was under the control of the ventricle specific myosin light chain-2 （mlc-2v）.Results: Our data demonstrated GFP was expressed in transfected neonate rat ventricular myocytes in vitro but not in transfected cell lines. Conclusion: the established viral vector AdmlcGFP allows heart specific expression of a foreign gene.