人GDNF基因的克隆

CLONING OF HUMAN GDNF GENE

目的：克隆可表达人胶质细胞源性神经营养因子的基因。方法：从脑胶质细胞瘤患者术后的脑瘤组织中提取总ＲＮＡ，以ＲＴＰＣＲ方法扩增出了一段约５５８ｂｐ的ｃＤＮＡ片段，将这一片段克隆于ｐＵＣ１８载体中，进行全序列分析。结果：证实该研究所克隆的ｃＤＮＡ是编码正确的人ＧＤＮＦ基因。与发表于ＧｅｎｅＢａｎｋ上的序列相比，发现该研究克隆的基因有一段７８ｂｐ的核苷酸序列缺失，但不影响密码子的阅读框。结论：克隆到了编码正确的人ＧＤＮＦ的ｃＤＮＡ全序列，对帕金森病基因治疗的基础研究及临床应用具有重要意义。

Objective: To obtain the human glial cell linederived neurotrophic factor gene, which will be used in gene therapy for human Parkinson disease. Methods: 200 mg of brain glioma tissues were used to extract total RNA. A 558bp positive cDNA band was obtained by RTPCR method. This fragment was cloned into the Hinc Ⅱ site of pUC18 vector. The cloned fragment was sequenced. Results: Compared with the sequence deposited in GeneBank(L19036,L15306),a 78bp fragments deletion was found , but the reading frame was unchanged, and the sequence correct. Conclusion: The cloned human GDNF gene will play an important role in gene therapy research and its clinical utility in the future for Parkinson disease.