质粒型单纯疱疹病毒载体介导GDNF和GFP基因在体外培养的BHK细胞和大鼠脊髓、背根节神经元中的转移和表达

为了观察质粒型单纯疱疹病毒载体介导的外源性胶质细胞源性神经营养因子和绿色荧光蛋白基因在体外培养的金黄地鼠肾细胞以及脊髓神经元和背根节神经元中的转移和表达 ,本研究采用了以质粒型单纯疱疹病毒载体为基础构建的分别含有重组胶质细胞源性神经营养因子和绿色荧光蛋白基因的混合毒株 dv HSV-GDNF和 dv HSV-GFP感染体外培养的金黄地鼠肾细胞、脊髓神经元和背根节神经元。用免疫组织化学方法和荧光显微镜观测法分别检测了胶质细胞源性神经营养因子和绿色荧光蛋白基因的转移和表达。结果发现 :质粒型单纯疱疹病毒载体可以成功地将外源性胶质细胞源性神经营养因子和绿色荧光蛋白基因导入金黄地鼠的肾细胞以及脊髓神经元和背根节神经元中。提示质粒型单纯疱疹病毒载体可以作为转基因胶质细胞源性营养因子治疗脊髓损伤的转移载体 ,为脊髓损伤的基因治疗提供了实验基础。绿色荧光蛋白作为报告基因 ,也可因其适用广泛、观察简便等特性而得到更加广泛的应用。

THE EXPRESSION OF GDNF AND GFP MEDIATED THROUGH AMPLICON HSV BY BHK CELLS,NEURONS OF SPINAL CORD AND DRG OF RAT IN VITRO

In order to observe the gene transferring and expressing of foreign GDNF and GFP mediated through a amplicon herpes simplex virus (HSV) vector system by the neurons of spinal cord and dorsal root ganglia (DRG) in vitro. BHK cells, neurons of spinal cord and DRG were transfectted through amplicon HSV based mixed virus stock dvHSV GDNF and dvHSV GFP in vitro. And the transferring and expressing of GDNF and GFP gene with immunohistochemical and fluorescent techniques were tested. The results showed that the amplicon HSV vector system transferred GDNF and GFP gene into BHK cells, neurons of spinal cord and DRG. These suggested that the amplicon HSV vector system could be used as transfer vector for GDNF to help repair spinal cord injury. GFP, as a reporter gene with specific property such as species independent fashion and simple assay, could be widely used in molecular biology.