论文部分内容阅读
目的 探讨使用腺伴随病毒 (AAV)载体介导编码多巴胺合成相关的三种生物酶基因的共转移。方法 使用AAV载体将多巴胺生物合成相关的三种生物酶———酪氨酸氢化酶 (TH)、芳香族氨基酸脱酶 (AADC)、以及三磷酸鸟苷环水解酶Ⅰ (GGH)基因共转导入 2 93细胞 ,以Western印迹分析检测TH、AADC、GCH的表达 ,以高效液相色谱法检测细胞内多巴胺水平。结果 TH、AADC、GCH的可在使用三种AAV载体 (AAV TH、AAV AADC、AAV GCH)转导的细胞中有效共表达 ,进一步分析发现 ,这种共表达可使该细胞有效地自发生成多巴胺。结论 使用AAV载体进行TH、AADC、GCH三重基因共转导是有效的 ,这对帕金森病基因治疗的研究具有重要意义。
Objective To investigate the use of adeno-associated virus (AAV) vectors to mediate the co-transfer of three genes involved in dopamine biosynthesis. Methods AAV vectors were used to co-transfect three enzymes involved in dopamine biosynthesis: tyrosine hydrogenase (TH), aromatic amino acid dehydratase (AADC), and guanosine triphosphate cyclohydrolase I (GGH) 293 cells were transfected, the expression of TH, AADC and GCH was detected by Western blotting, and the level of intracellular dopamine was detected by high performance liquid chromatography. Results TH, AADC, GCH were efficiently co-expressed in cells transduced with three AAV vectors (AAV TH, AAV AADC, AAV GCH) and further analysis revealed that this co-expression allowed the cells to spontaneously produce dopamine . CONCLUSION: The co-transduction of TH, AADC and GCH genes with AAV vector is effective, which is of great significance for the study of gene therapy of Parkinson’s disease.