CRIF1基因启动子克隆及活性分析

Cloning and activity analysis of CRIF1 promoter

目的研究急性髓性白血病基因1(AML-1/RUNX1)调控CR6相互作用因子(CRIF1)基因启动子的主要活性区域。方法采用PCR技术克隆CRIF1基因启动子并测序,构建截短突变体,以双荧光素酶报告系统检测AML-1/RUNX1调节CRIF1基因启动子的主要活性区域。结果克隆出CRIF1基因启动子-2 505—+9共计2 514 bp的活性区域,并构建了截短突变体CRIF1-1 800、1 500、1 000、600及300 bp;较之CRIF1基因启动子全长序列转录活性,CRIF1-300 bp突变启动子活力明显下降,CRIF1-600 bp内启动子活力与其相当,CRIF1-1 500 bp区启动子活性是它的2.8倍。结论本研究提示CRIF1基因启动子-1 500—-2 514 bp间可能是AML-1/RUNX1对其进行转录负性调控的主要活性区域。

Objective To analyze the main active region in the promoter of CRIF1 regulated by AML-1/RUNX1. Methods The 2 514 bp fragment of CRIF1 promotor was cloned and truncated mutants （ CRIF1 - 1 800 bp, - 1 500 bp, - 1 000 bp, -600 bp, -300 bp） were constructed. Compared with the full -length CRIF1, the transcriptional activity of promotor decreased when it had been truncated in - 300 bp, equally unchanged in - 600 bp and 2. 8 times in - 1 500 bp. Conclusion The study displays that the main active region in promotor of CR/F1 regulated by AML-1/RUNX1 was falls in - 1 500-- -2 514 bp.