摘要
旨在探明坝上长尾鸡小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)基因启动子活性区与结构,为研究MITF基因在毛囊组织中的表达调控提供依据。本研究采用双荧光素酶表达载体的方法,构建了6个含有不同长度坝上长尾鸡MITF基因启动子片段的pGL3-Basic表达载体及4个核心启动子区突变载体,转染DF1细胞48h后检测双荧光素酶活性。结果,确定了鸡MITF基因启动子的核心区域为-660^+200bp。其中突变位点-579bp、-505bp、-274bp、-220bp、-203^-202bp、-98bp、-46bp、-14bp、+1bp、+28bp对MITF基因启动子活性有较大影响,在突变区域预测到HOX家族(HOXA3、HOXD8、HOXD9、HOXD10、HOX11)、NF-1、DBP、TFIID和TFIIB与MITF序列的结合性发生了变化。
The research was designed to study the activity and the structure of MITFpromoter and to provide clues for studying this gene＇s regulation mechanism in hair follicle.Dual-luciferase expression vectors were constructed and transfected to DF1 cells with lip2000.The dual-luciferase detection kit was used to measure the relative luciferase activity.The 6 expression vectors with different promoter regions of MITFgene and 4 mutant vectors of the core promoter region were constructed in Bashang Long-tail chicken.The core promoter region from-660 bp to＋200 bp was identified in chicken MITFgene.The function of-579 bp,-505 bp,-274 bp,-220 bp,-203--202 bp,-98 bp,-46 bp,-14 bp,＋1 bp and＋28 bp sites in regulating promoter activity was critical.The similarities of transcription factor binding sites including HOX family（HOXA3,HOXD8,HOXD9,HOXD10 and HOX11）,NF-1,DBP,TFIID and TFIIB to MITF were changed predicted by software.
作者
刘小辉
彭永东
周荣艳
张传生
李祥龙
LIU Xiao-hui;PENG Yong-dong;ZHOU Rong-yan;ZHANG Chuan-sheng;LI Xiang-long(College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China;College of Animal Science and Technology, Hebei Normal University of Science ＆ Technology, Qinhuangdao 066004, China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2018年第6期1116-1123,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
河北省蛋鸡体系产业技术创新团队(HBCT2013090206)
河北省科技计划项目(15226302D)
关键词
坝上长尾鸡
MITF基因
启动子
转录因子
Bashang Long-tail chicken
MITF gene
promoter
transcription factor
作者简介
刘小辉（1988-），女，河北邯郸人，博士生，主要从事动物遗传育种研究，E-mail：liuxiaohui1000@163．com;通信作者：李祥龙，教授，E-mail：lixianglongcn@yahoo．com
