人硫氧还蛋白cDNA的克隆及序列测定

Cloning and sequencing of the cDNA for human thioredoxin

目的 克隆人硫氧还蛋白 (HumanThioredoxin ,hTRX)cDNA序列并进行序列测定。方法 应用RT PCR技术 ,以 1 43(TK- )人骨肉瘤细胞RNA为模板 ,扩增出hTRX成熟蛋白的cDNA基因 ,并克隆至 pGEM TEasy载体中进行基因序列测定。结果 将所得序列与GenBanK(J0 4 0 2 6)提供的序列比较 ,其酶活性中心 (Trp Cys Gly Pro Cys)和基序与已知序列一致 ,第 1 80、2 84位碱基与已知序列不同 ,编码的氨基酸也发生了改变。结论 克隆得编码hTRX成熟蛋白的cDNA基因 ,为进一步探讨hTRX的生物活性和应用奠定了基础。

Objective Cloning and sequencing of the cDNA for human thioredoxin. Methods The cDNA of human thioredoxin was obtained by RT PCR from 143(TK -) human osteosarcoma cell. The amplification products were then cloned into pGEM T Easy Vector and sequenced.Results By comparing the cDNA sequence we get with that of the GenBank(J04026),we found that the conserved enzymatic active site(Trp Cys Cly Pro Cys) of the two was identical with each other. However,there were two differences at bp180 and bp284 and the counterpart amino acids were also changed.Conclusion Recombinant human thioredoxin(hTRX) gene clone was established for further study on biological functions and utilization of hTRX.