利用荧光素酶报告基因法研究转录因子USF1结合调控马鹿MicroRNA PC-5p-4811基因启动子机制  

The Mechanism on Transcription Factor USF1 to Regulate MicroRNA PC-5p-4811 Gene Promoter of the Red Deer Based on the Luciferase Reporter Method

作　　者：高仰 韩青 吴玄烨 索婧媛 金庆梅 刘学东[1] 郑冬[1] GAO Yang;HAN Qing;WU Xuanye;SUO Jingyuan;JIN Qingmei;LIU Xuedong;ZHENG Dong(College of Wildlife and Protected Area,Northeast Forestry University,Harbin,150040,China)

机构地区：[1]东北林业大学野生动物与自然保护地学院,哈尔滨150040

出　　处：《野生动物学报》2025年第1期63-71,共9页CHINESE JOURNAL OF WILDLIFE

基　　金：国家自然科学基金面上项目(31671283)。

摘　　要：为了探究转录因子上游刺激因子1(USF1)调控马鹿(Cervus elaphus)microRNA PC-5p-4811(miR-4811)基因转录的机理,综合运用核心启动子和转录因子结合位点生物信息学预测分析、基因克隆、点突变和双荧光素酶报告基因法等技术研究USF1结合miR-4811启动子分子机制和功能。结果显示:(1)截切突变证实马鹿miR-4811基因的核心启动子位于上游-3174~-2966 bp;(2)转录因子USF1在-3378~+73 bp区域共有2个DNA结合位点(-3046~-3031 bp、-3045~-3018 bp),均位于miR-4811核心启动子内;(3)USF1作为激活因子可直接结合上述位点激活miR-4811核心启动子上调双荧光素酶报告基因转录表达。研究结果揭示了USF1参与调节microRNA基因转录表达的机理,并为后续USF1/miR-4811调节轴参与调控鹿茸再生发育机制提供了重要基础数据。To explore mechanism of the upstream stimulatory factor 1(USF1)regulating transcription of microRNA PC-5p-4811(miR-4811)gene in red deer(Cervus elaphus),we employed a series of methods,including bioinformatic prediction for core promoters and transcription factor binding motifs,gene cloning,site-directed mutagenesis,and dual-luciferase re⁃porter assays.The results showed:(1)Truncation assays confirmed that core promoter of miR-4811 gene was located within the upstream region from-3174 bp to-2966 bp.(2)USF1 owned two DNA binding sites(-3046 bp to-3031 bp,and-3045 bp to-3018 bp),both located within the core promoter region.(3)USF1,as an activator,could bind to the sites directly,activate the core promoter of miR-4811 and further upregulate the dual-luciferase reporter gene expression.The study provided the basic data to reveal the mechanism on USF1 regulating microRNA gene transcription,and would of⁃fer essential data for USF1/miR-4811 axis controlling antler regeneration and development in the future.

关 键 词：USF1 马鹿 MicroRNA PC-5p-4811 启动子 转录调控 

分 类 号：Q78[生物学—分子生物学]