Cheng Saiyu, Ruan Huaizhen, Yang Zhong, Wu Xigui
Department of Neurobiology, The Third Military Medical University, Chongqing Institute of Neuroscience, Chongqing 400038, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2008 Jun;25(3):642-6.
By genetic recombinant technique, the rat GDNF cDNA was recombinated to the retroviral vector pLXSN. The recombinant plasmid pLXSN-GDNF was verified by digestion with restriction endonucleases and PCR. Then neural stem cells (NSCs) were infected with pLXSN-GDNF. Immunocytochemistry, RT-PCR and western-blot were used to detect the transfection effect. Results showed that GDNF cDNA was cloned into retroviral vector pLXSN correctly, and the pLXSN-GDNF can infect NSCs efficiently. These results provide the possibility for transplantation and gene therapy with GDNF of nervous system diseases and injury.
通过基因重组技术，将大鼠胶质细胞源性神经营养因子（GDNF）cDNA重组到逆转录病毒载体pLXSN中。重组质粒pLXSN-GDNF经限制性内切酶酶切和聚合酶链反应（PCR）验证。然后用pLXSN-GDNF感染神经干细胞（NSCs）。采用免疫细胞化学、逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测转染效果。结果表明，GDNF cDNA正确克隆到逆转录病毒载体pLXSN中，且pLXSN-GDNF能有效感染NSCs。这些结果为GDNF用于神经系统疾病和损伤的移植及基因治疗提供了可能性。
