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当前位置: 首页 > 文章 > 北极狐TYRP1基因启动子活性及转录调控区域分析 中国兽医学报 2018,38 (12) 2366-2373
Position: Home > Articles > Promoter and transcription regulatory region in fox TYRP1 gene Chinese Journal of Veterinary Science 2018,38 (12) 2366-2373

北极狐TYRP1基因启动子活性及转录调控区域分析

作  者：
郑晓宁;王瑞宁;王亚琪;郭敏;巩元芳;刘铮铸;彭永东;李祥龙
单  位：
河北农业大学动物科技学院;河北科技师范学院动物科技学院
关键词：
北极狐;TYRP1基因;启动子;点突变;转录调控
摘  要：
通过分析调控北极狐毛色基因TYRP1启动子核心区域及转录因子,为探究该基因的表达调控机制提供理论依据,并为狐狸毛皮品质分子育种和彩色毛皮新材料的创制提供思路。通过基因组测序技术获得了北极狐TYRP1基因启动子序列,并利用生物信息学方法对北极狐TYRP1基因核心启动子区域和转录因子结合位点进行预测;以北极狐基因组DNA为模板,PCR扩增北极狐TYRP1基因不同长度的启动子缺失片段克隆至pGL3-Basic载体,将重组质粒瞬时转染到A375和293T细胞,利用双荧光素酶基因检测仪进行活性验证。结果表明,成功构建了9个含有不同长度启动子片段的重组质粒,经双荧光素酶活性检测发现北极狐TYRP1基因-699/+35区域为核心启动子区域,-699/-93区域存在着TYRP1基因正调控元件。生物信息学预测分析发现该区域存在4个转录因子结合位点;利用重叠延伸PCR技术成功构建了4个突变载体,经双荧光素酶活性检测发现4个突变载体活性均显著下降(P<0.05),表明这4个转录因子是北极狐TYRP1基因转录调控的正调控元件。本研究确定了北极狐TYRP1基因启动子核心区域-699/+35,Sp1(-656/-646)、CREB(-598/-589)、Sp1(-539/-530)和Sp1(-163/-154)为北极狐TYRP1基因转录的正调控元件。
译  名：
Promoter and transcription regulatory region in fox TYRP1 gene
作  者：
ZHENG Xiao-ning;WANG Rui-ning;WANG Ya-qi;GUO Min;GONG Yuan-fang;LIU Zheng-zhu;PENG Yong-dong;LI Xiang-long;College of Animal Science and Technology,Hebei Normal University of Science & Technology;College of Animal Science and Technology,Hebei Agricultural University;
关键词：
fox;;TYRP1 gene;;promoter;;point mutation;;transcriptional regulation
摘  要：
The research was designed to investigate the activity region and transcription factors of fox TYRP1 gene,and to provide a theoretical reference for exploring the breeding of fox and creating the new fur materials.The gene promoter sequence of fox TYRP1 gene was obtained through transcriptome sequencing technology,and the method of bioinformatics was used to predict the core promoter region of TYRP1 gene and the transcription factor binding site.The fragment in a 5′flanking region was amplified and cloned into the vector pGL3-Basic.The positive colonies were identified and sequenced.The recombinant plasmid was transfected to A375 and 293 Tcells,and the activity was verified by the dual-luciferase assays system.The results showed that 9 fragments with different lengths of promoter regions were amplified and cloned into the vector pGL3-Basic.The region-699/+35 of fox TYRP1 gene was detected as the core promoter region by dual-luciferase assays system.There were some positive regulatory elements in the region from-699/-93.The bioinformatics prediction analysis revealed that there were 4 transcription factor binding sites in the region.Using the overlap extension PCR technology successfully constructed 4 mutation vectors.Their activity were significantly decreased by dual luciferase assay.It is suggests that these 4 transcription factors were the positive regulatory elements in fox TYRP1 gene transcription.In this study,the identified core regions of TYRP1 gene promoter is-699/+35.Sp1(-656/-646),CREB(-598/-589),Sp1(-539/-530)and Sp1(-163/-154)binding sites are the positive regulatory elements of fox TYRP1 gene.

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