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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 To clone cDNA sequence of human Thioredoxin (hTRX) and determine its sequence. Methods RT - PCR was used to amplify the cDNA of hTRX mature protein using human osteosarcoma cell line 1 43 (TK -) as a template and cloned into pGEM TEasy vector for sequencing. Results The obtained sequence was compared with the sequence provided by GenBanK (J0 4026). The Trp Cys Gly Pro Cys and its motif were consistent with the known sequences. Different sequences, encoded amino acids have also changed. Conclusion The cloned cDNA encoding hTRX mature protein laid the foundation for further study on the biological activity of hTRX and its application.