RLM-RACE法确定胰岛素降解酶基因的转录起始位点

摘要

目的 精确定位胰岛素降解酶基因的转录起始位点,为今后研究该基因启动子区的转录调控及其参与疾病的发病机制奠定基础.方法采用5' cDNA末端快速扩增技术,得到多个胰岛素降解酶基因cDNA 5' 末端序列,连接至PUC19 T载体中,鉴定阳性克隆并测序.结果 根据两次实验结果,确认胰岛素降解酶基因的5' UTR序列长度为32bp.结论胰岛素降解酶基因存在单一的转录起始位点,位于翻译起始点上游32bp处,碱基为G.%Objective To determine the transcription start site of insulin degrading enzyme gene for further study on the transcriptional regulation in promoter region and the pathogenesis. Methods We used 5' rapid amplification of cDNA ends to obtain insulin-degrading enzyme gene 5 'cDNA ends, and connected it to the PUC19 T vector, then the positive clones were identified and sequenced. Results According to the repeated experimental results, we confirmed that the 5'UTR of insulin degrading enzyme gene was 32bp. Conclusion The transcription start site of insulin-degrading enzyme gene is located 32 bp upstream of the translation start codon, the base is G.