心肌特异性表达腺病毒载体的构建及鉴定

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源基因在心肌细胞内特异性表达。

The construction and identification of cardic specific expression adenovirus vector

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.