畜牧与兽医2018，Vol.50Issue(6)：12-18,7.
猪速激肽3基因启动子活性及其表达调控的初步研究
Analysis of the promoter activity and its regulation of TAC3 gene expression in pig
摘要
Abstract
This study was to determine the sequence of the core promoter of tachykinin 3 gene (TAC3),and to explore the role of transcription factors in regulation of the TAC3 core promoter region.The deletion fragments of 5'end of TAC3 were amplified by PCR,taking the porcine ear genomic DNA as a model.The fragments were cloned into the pGL3-basic vector to build gene promoter recombinant plasmids,and then were transfected into porcine ovarian granulosa cells.The core promoter region was determined by double luciferase activity analysis.Chromatin immunoprecipitation assay (CHIP) was performed to determine if the transcription factors physically bound to the promoters of TAC3.The transcription factor overexpression vector and the small interfering RNA (siRNA) transfected porcine ovarian granulosa cells were formed,and qRT-PCR was used to detect changes in expression of TAC3 mRNA.The results showed that the deletion fragments of the 5'end of TAC3 were successfully constructed.The double-luciferase reporter system showed that the core promoter region of TAC3 was at-1 310/ -544.The-2 486/-1 633 region might contain negative regulation elements,and the-1 310/-544 region might contain positive control elements.ChIP assay showed that C/EBPβ and YYl bound to the-653/-639 and-873/-856 regions of TAC3,respectively.Overexpression of C/EBPβ and YY1 enhanced the activity of the TAC3 gene promoter (P＜0.05) and raised the mRNA expression level (P＜0.01).Disturbing the expression of C/EBPβ reduced the expression of mRNA of TAC3 (P＜0.01);and disturbing the expression of YY1 also reduced the gene promoter activity (P＜0.05).The findings here suggested that the C/EBPβ and YY1 genes acted on the core promoter region of TAC3,and contributed to the transcriptional activity of the gene.关键词
TAC3基因/5'端缺失片段/C/EBPβ/YY1/猪Key words
TAC3 gene/deletion fragments/C/EBPβ/YY1/pigs分类
农业科技引用本文复制引用
李忠慧,袁晓龙,邢燕,林长光,张哲,张豪,李加琪..猪速激肽3基因启动子活性及其表达调控的初步研究[J].畜牧与兽医,2018,50(6):12-18,7.基金项目
国家现代农业产业技术体系(CARS-35) （CARS-35）
福建省科技重大专项(2012NZ0003-3) （2012NZ0003-3）
国家科技基础性工作专项(2014FY120800) （2014FY120800）
