人甲胎蛋白转录调控序列的克隆

Cloning of Transcriptional Regulatory Sequences in Human α-Fetoprotein Gene

本文根据已知的甲胎蛋白基因5’-端增强子调控顺序,自行设计一对引物(引物1∶5’-TGCAAGCTTATGATTCCCAAATATC-3’引物2∶5’-GTCGAATTCGTGGCCTGGATAAAGCTGAGT-3’),用PCR方法从染色体DNA中成功地克隆416bp长度的甲胎蛋白转录调控序列(AFPTRS),经限制性内切酶鉴定及DNA序列分析证实与报道的顺序基本一致。可望应用AFPTRS调控细胞因子在肝癌细胞中高效特异地表达。

The 5'-flanking region of the human α-fetoprotein(AFP) gene contains transcriptional regulatory sequences(TRSs) which up-regulate AFP gene expression with cell-specific enhancer activity in hepatoma cells. A couple of primers(Primerl: 5'-TGCAAGCTTATGATTCCCAAATATC-3'; Primer 2: 5'-GTCGAATTCGTGGCCTGGA TAAAGCTGAGT-3') were designed for synthesis and purification according to the known sequences of 5'-flanking region. AFPTRSs of 416 base pairs were amplified from human chromosome DNA by PCR, The identification of the AFPTRSs was confirmed to be consistent with reported sequences. The AFPTRSs can be applied to regulating the specific expression of cytokines in hepatoma cells.