人δ珠蛋白基因启动子区CAAT盒C→T突变对其转录的影响

The Transcriptional Regulation Study on Human δ Globin Gene with CAAT Box C→T Point Mutation in Its Promoter

目的研究人δ珠蛋白基因启动子区-64位C→T点突变对其转录调控的影响。方法将人δ基因启动子区-64位C突变为T后,克隆于去除强启动子CMV的真核基因表达载体pcDNA3.1穴-雪/Myc-HisA中,分别将含野生型CAAT盒和突变型CAAT盒的人δ基因转染鼠红白血病(MEL)细胞,经DMSO诱导细胞分化,利用半定量RT-PCR技术检测人δ基因转录的变化。结果δ基因启动子区CAAT盒C→T点突变后,含突变型CAAT盒的人δ基因的转录水平是含野生型CAAT盒的2.2倍。结论在mRNA水平直接证实δ基因启动子区CAAT盒的缺陷是影响其转录的重要原因。

Objective To study the transcriptional regulation of humanδglobin gene with C→T point mutation at-64in its promoter. Methods Humanδglobin genes including wild CAAT box and mut-ant CAAT box(-64C→T)were separately cloned into eukaryotic expression vector pcDNA3.1(-)/Myc-His A which was cut out the strong promoter CMV,transfected MEL cells,and induced by DMSO to express.The transcriptional regulation of humanδglobin gene was analysed using semi-quan-titative RT-PCR. Results The expression level of humanδglobin gene with mutant CAAT box was2.2-fold as high as that with wild CAAT box. Conclusion The defective CAAT box of humanδglobin gene promoter region may be one of the major reasons for its low expression level.