人NRAGE基因启动子转录活性的初步研究

Transcription Activity of Human NRAGE Gene Promoter

利用生物信息学软件搜寻NRAGE启动子区域的转录因子结合位点,并根据这些位点利用PCR技术克隆NRAGE启动子不同区域的缺失突变体,分别插入Luciferase报告载体pGL3-Basic载体中,构成8种缺失突变体的报告基因表达载体.通过双荧光素酶体系检测在HEK-293细胞中NRAGE启动子不同区域的转录活性,从而确定起主要作用的启动子区域.结果发现NRAGE基因转录起始位点上游-300 bp到-100 bp的区域起到主要的转录调控作用.

We analyzed human NRAGE promoter region by bioinformatic software and found putative binding sites of transcription factors, then 8 different fragments were amplified by PCR and the mutant promoters were inserted to upstream of the luciferase gene in luciferase report vector pGL3-Basic and detected the transcription activity by dual-luciferase reporter assay systerm. At transcription level we detected that transcription enhancer elements may exist in the region - 300 bp to - 100 bp.