人神经营养素-3全长基因克隆及序列分析

Cloning and sequencing of human neurotrophin-3 intact gene

目的 :扩增人神经营养素 3(humanneurotrophin 3,hNT 3)全长基因并进行序列分析。 方法 :应用聚合酶链反应技术 ,以一个健康成人基因组DNA为模板 ,扩增出编码hNT 3的全长基因 ,并克隆至pMD18 T载体中进行基因测序。结果 :所得序列与Genbank提供的序列 (M 6 1180 )对比显示 ,除hNT 3中第 5 5 5位碱基由C→T外 ,余序列同已知序列完全一致 ,改变的碱基并不影响基因编码的氨基酸序列。结论 :成功地获得了hNT 3的全长基因 ,且序列与已知序列高度一致。

Objective:To amplify and sequence the human neurotrophin-3(hNT-3) intact gene.Methods:hNT-3 intact gene was obtained by polymerase chain reaction using human genomic DNA as a template and cloned into pMD18-T vector and sequenced.Results:DNA sequence comparison of the cloned fragments and the reported hNT-3(M61180) demonstrated that the gene sequence was identical to that had been reported before except one base(555th) changing from C to T,but it had no influence on the encoded amino acids.Conclusions:hNT-3 intact gene was obtained and its sequence was highly consistent with ones reported.