人脑源性神经营养因子基因克隆及序列分析

目的　克隆人脑源性神经营养因子 (hBDNF)基因并进行序列分析。方法　提取健康成人末梢血白细胞基因组DNA作为模板 ,应用PCR技术和T 载体克隆法克隆hBDNF基因 ,筛选阳性克隆、酶切鉴定 ,并进行序列测定和分析。结果　DNA序列测定的结果与GenBank提供的已知序列 (M6 1 1 81 )比较 ,所克隆的hBDNF基因从起始密码子ATG到终止密码子TAG全长共744bp ,序列完全相同。结论　自人基因组DNA中克隆hBDNF基因 ,为进一步开展阿尔茨海默病 (AD)的基因治疗积累了资料。

Molecular cloning and sequencing of the human brain-derived neurotrophic factor gene

Objective Molecular cloning and sequencing of the human brain-derived neurotrophic factor(hBDNF) gene. Methods Extracting the human genomic DNA from the white blood cells as templates,the hBDNF gene was cloned by using PCR and T-vector cloning method.The positive clone was screened and identified by the restriction enzymes,and then the cloned amplified fragment was sequenced and analyzed.Results Compared the cloned hBDNF gene with the GenBank(M61181) sequence,we demonstrated that both of sequence were from the initiation codon ATG to the termination TAG identically. The cloned hBDNF gene was 744bp length.Conclusion Cloning the hBDNF gene from the human genomic DNA has paved the way for further study on gene therapy of Alzheimer's Disease(AD).