The research was designed to study the activity and thestructure of MITF promoterand to provide clues for studying this gene's regulation mechanism in hair follicle.Dual-luciferase ex-pression 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 MITF gene 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 MITF gene.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 ac-tivity 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.%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48 h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～+200 bp.其中突变位点－579 bp 、－505 bp 、－274 bp 、－220 bp 、－203～－202 bp 、－98 bp 、－46 bp 、－14 bp 、+1 bp 、+28 bp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.
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