GFP标记的GDNF重组腺病毒载体的构建和体内外表达

Construction of a GFP-labeled Recombinant Adenovirus Harbouring GDNF and Its Expression in Rat Neural Cells in vitro and in vivo

通过构建绿色荧光蛋白(GFP)和胶质细胞源性神经营养因子(GDNF)共表达的腺病毒载体,讨论其在治疗脊髓和脑有关神经系统疾病的潜在应用价值,了解腺病毒载体在大鼠体内的分布及在体外分泌外源基因产物能力,并对其生物安全性作了初步考察,为GDNF重组腺病毒载体在临床和基础研究中的应用作了一定准备.

A recombinant adenoviral vector harbouring and coexpressing green fluorescent protein (GFP) and glial cell linederived neurotrophic factor (GDNF) was constructed. The construct AdGDNFEGFP was microinjected into the cortex cerebri,corpus striatum,lumbus segment of medulla spinalis,cisternae subarachnoideales and substantia grisea of medulla spinalis of rats. Then the distribution and expression of the exogenous genes in different types of neural cells were observed under fluorescent microscope by using GFP as an indicator. The recombinant virus was used to transfect rat/mouse cell lines NSC and NIH3T3 cells in vitro. The bioengineered cell line coexpressing GDNF and GFP was preliminarily abtained. According to the experimented results,the biosafety and the prospective of applying the adenovector thus constructed to clinical treatment of nervous system diseases were discussed.