人神经生长因子β亚基cDNA的克隆及序列分析

Cloning and sequencing of human β nerve growth factor cDNA gene

目的：对国人神经生长因子β亚基（βＮＧＦ）ｃＤＮＡ 进行克隆及序列分析，为研究βＮＧＦ 的生理功能及其在临床应用提供基础与手段。方法：从人海马组织分离，纯化总ＲＮＡ，直接进行逆转录，用ＰＣＲ 扩增βＮＧＦ 的ｃＤＮＡ 得到６５０ｂｐ 的ＤＮＡ 片段，利用Ｔ－ Ａ 克隆载体法，制备重组质粒，克隆βＮＧＦｃＤＮＡ。结果：βＮＧＦ 的ｃＤＮＡ 全序列１０７４ ｂｐ ，其蛋白编码为６３０ ｂｐ ，由２１２ 个氨基酸组成，Ｎ 端为信号肽，中间区为前肽，Ｃ 端为成熟肽。结论：国人βＮＧＦ 序列分析结果与ｇｅｎｅ ｂａｎｋ 中的βＮＧＦ 序列完全一致。

AIM:Cloning and sequencing the nerve growth factor β subunit (βNGF) cDNA of Chinese to study deeply on NGF the physiological function and clinical uses. METHODS:Total RNA was obtained and purified from embryo hippocampus, then reverse transcription and PCR were utilized to get the 650 bp cDNA of βNGF. Using A-T vector PCR Ⅱ, we got recombinant βNGF cDNA PCRⅡ.RESULTS:The βNGF cDNA a 1 074 bp sequence, 636 bps of this cDNA coding 212 amino acid protein-βNGF. The N terminal of the protein was signed peptide, the middle region, propeptide, and the C terminal, mature peptide. CONCLUSION:The βNGF cDNA sequence of Chinese people was completely conformed with what was published in gene bank. So there were no difference in βNGF cDNA between Chinese and foreigners.