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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、观察简便等特性而得到更加广泛的应用
In order to further explore new ways of treating central nervous system injury, we constructed a recombinant plasmid containing glial cell line-derived neurotrophic factor (GDNF) and green fluorescent protein (GFP) gene using plasmid vector herpes simplex virus (H SV) The recombinant vectors H SV - GDNF and H SV - GFP were used to infect cultured BHK cells and primary cultured rat skeletal muscle cells with the packaged mixed strains respectively. After 2 d, G DNF immunohistochemistry was performed on cells infected with HSV - G DNF; cells infected with HSV - GFP were observed under a fluorescence microscope. As a result, it was found that plasmid-type H SV vectors successfully introduced exogenous GDN F and GFP genes into BH K cells and primary cultured rat skeletal muscle cells. Tip: Plasmid H SV vector can be used as a transgene GDN F in the treatment of central nervous system injury, which has broad prospects for the treatment of central nervous system injury. GFP as a reporter gene, but also because of its broad spectrum of applications, observation and other characteristics of a more widely used