牛TARDBP基因核心启动子鉴定与转录调控分析OA北大核心CSTPCD

[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.