质粒型HSV载体介导的GDNF和GFP基因在BHK细胞和骨骼肌细胞中的转移和表达

THE EXPRESSION OF GDNF AND GFP GENE MEDIATED THROUGH AMPLICON HSV BY BHK CELLS AND SKELETAL MUSCLE CELLS IN VITRO

为了进一步探索治疗中枢神经损伤的新途径 ,本实验以质粒型单纯疱疹病毒 ( H SV)为载体 ,分别构建了含有胶质细胞源性神经营养因子 ( GDN F)和绿色荧光蛋白 ( GFP)基因的重组载体 H SV - GDNF和 H SV- GFP,用包装出的混合毒株分别感染体外培养的 BHK细胞及原代培养的大鼠骨骼肌细胞。 2 d后 ,对感染 HSV - G DNF组的细胞进行 G DNF免疫组织化学反应 ;感染HSV- GFP组的细胞在荧光显微镜下进行观察。结果发现 :质粒型 H SV载体可成功地将外源性 GDN F和 GFP基因导入 BH K细胞和原代培养的大鼠骨骼肌细胞。提示 :质粒型 H SV载体可以作为转基因 GDN F治疗中枢神经系统损伤的转移载体 ,为中枢神经系统损伤的治疗展示了广阔的前景。 GFP作为报告基因 ,也可因其适用广谱。

In order to explore a new way for GDNF to cure the injured CNS, the present study transferred the recombinant GDNF gene and green fluorescent protein (GFP) gene (control) into BHK cells (baby hamster kidney cells) and skeletal muscle cells of rat in vitro through an amplicon herpes simplex virus (HSV) vector system. With immunohistochemical staining technique and fluorescent microscope, we testified tesified that these two defective virus dvHSV GDNF and dvHSV GFP can effectively transfer gene GDNF and GFP into BHK cells and skeletal muscle cells of rat in vitro. These results suggested that the amplicon HSV vector system could be used as effective tranfer vector for GDNF to treat CNS injury. GFP, as a reporter gene with specific property such as species independent fashion and simple to use, will be widely used in molecular biology. (Figures 1～4 on plate 22)