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目的 :建立简便快速地获取人胶质细胞源神经营养因子 ( GDNF)基因的 PCR方法。方法 :根据从Gene Bank中调出的人 GDNF基因全序列设计引物 ,提取人基因组 DNA做模板行 PCR,经酶切和凝胶电泳鉴定产物。结果 :扩增出 4 2 5bp的人 GDNF基因 ,经 Eco R I酶切可见预期的 34 1 bp片段 ,证实获得正确的产物。结论 :成功建立了一种简便快速地从人基因组 DNA获取人 GDNF基因的 PCR方法 ,避免了其他作者为获得 GDNF基因而采取收集神经胶质瘤标本 ,提取 RNA,逆转录 PCR等较为繁琐的步骤 ,省时省力 ,经济实用。
Objective: To establish a simple and rapid PCR method for obtaining human glial cell line-derived neurotrophic factor (GDNF) gene. Methods: According to the complete sequence of human GDNF gene from Gene Bank, primers were designed and human genomic DNA was extracted for PCR. The products were identified by restriction enzyme digestion and gel electrophoresis. RESULTS: A 4 2 5 bp human GDNF gene was amplified and the expected 34 1 bp fragment was digested with EcoRI to verify that the correct product was obtained. CONCLUSION: A simple and rapid PCR method for obtaining human GDNF gene from human genomic DNA has been successfully established, which avoids the cumbersome steps of other authors in collecting glioma specimens, RNA extraction and reverse transcription PCR in order to obtain GDNF gene , Save time and labor, economical and practical.