生物技术通报2024，Vol.40Issue(4)：306-318,13.DOI:10.13560/j.cnki.biotech.bull.1985.2023-1048
牛TARDBP基因核心启动子鉴定与转录调控分析
Identification and Transcriptional Regulation Analysis of Core Promoter in Bovine TARDBP Gene
摘要
Abstract
[Objective]The aim of this study is to analyze the transcriptional regulation mechanism of bovine TARDBP gene,as well as its structure,function and promoter active region.[Method]The complete coding region of bovine TARDBP gene was cloned by PCR.Bioinformatics software was applied to analyze the properties and the structure of bovine TARDBP protein.The expression of TARDBP gene was profiled in different tissues of lactating Holstein cows by real-time fluorescence quantitative PCR(RT-qPCR).The transcriptional initiation site of TARDBP gene was determined by 5'RACE technique,and the promoter region and 5'terminal deletion fragment of this gene were cloned by PCR.The CpG island and potential transcription factor binding site within the promoter region were predicted by bioinformatics software.The core promoter region was then confirmed by the dual-luciferase reporter system.[Result]Bovine TARDBP gene was expressed in multiple tested tissues including mammary gland.TARDBP protein was water-soluble,and it contained RNA recognition motifs and DNA binding sites.There were two CpG islands and a large number of transcription factor binding sites including SP1,PPARA,PPARD,PPARG,SREBF1 and so on in the promoter region.The results of dual-luciferase activity analysis showed that the core promoter region of bovine TARDBP was between-476 and-149,and Sp1,PPARG,PPARD and SREBF1 binding sites were found in the core promoter,but there was no TATA-box.[Conclusion]Bovine TARDBP gene is distributed in various tissues of dairy cows.The core promoter region of bovine TARDBP gene is positioned at-476 to-149,and there are transcription factor binding sites related to milk fat synthesis and secretion in this region.关键词
荷斯坦奶牛/TARDBP基因/生物信息学/启动子Key words
Holstein dairy cows/TARDBP gene/bioinformatics/promoter引用本文复制引用
张清兰,王洪程,张亚冉,鞠志花,王秀革,肖遥,王金鹏,魏晓超,高亚平,白福恒..牛TARDBP基因核心启动子鉴定与转录调控分析[J].生物技术通报,2024,40(4):306-318,13.基金项目
国家自然科学基金项目(32202648),山东省重点研发计划(竞争性创新平台)项目(2022CXPT010),山东省农业科学院农业科技创新工程(CXGC2023F10,CXGC2023A23) （32202648）
