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.

高 仰; 韩 青; 吴玄烨; 索婧媛; 金庆梅; 刘学东; 郑 冬

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 reporter assays. The results showed: (1) Truncation assays confirmed that core promoter of miR-4811 gene was located within the upstream region from -3 174 bp to -2 966 bp. (2) USF1 owned two DNA binding sites (-3 046 bp to -3 031 bp, and -3 045 bp to -3 018 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 offer essential data for USF1/miR-4811 axis controlling antler regeneration and development in the future.

TRANSCRIPTION factors; MOLECULAR cloning; RED deer; GENE expression; BINDING sites

Chinese Journal of Wildlife / Yesheng Dongwu Xuebao, 2025, Vol 46, Issue 1, p63

2310-1490

Academic Journal

10.12375/ysdwxb.20250107