To explore the activity of the pmel core promoter of Bashang long-tail chickens, we constructed dual-luciferase expression vectors and transiently transfected into DF1 cells with Lipofectamine 2000. We measured the luciferase activity with the dual-luciferase detection kit. The 1 268 bp fragment in 5-flanking region of the pmel gene in Bashang long-tail chickens was cloned. The region from -1 200 bp to +68 bp included 2 CpG islands and multiple transcription factor binding sites. We constructed 9 expression vectors with different promoter regions and a mutant vector of the core promoter region of the pmel gene of Bashang long-tail chickens. The core promoter region from -840 bp to +68 bp was identified in the pmel gene. The region from -590 to -525 bp negatively regulated the pmel gene during the transcription process. The -840--590 bp and -525--266 bp regions were positive regulatory regions. The polymorphic sites (-456, -435, -410, -374 and -341) had a significant effect on the promoter activity of the pmel gene.
为探明坝上长尾鸡的前黑素小体蛋白 (Pre-melanosomal protein，Pmel)基因核心启动子区，首先构建了双荧光素酶表达载体，通过脂质体瞬时转染鸡胚成纤维细胞DF1，并利用双荧光素酶检测试剂盒进行启动活性检测。成功克隆了坝上长尾鸡pmel 基因5侧翼区片段1 268 bp，预测启动子区 (−1 200–+68) 含有2 个CpG 岛和多种转录因子结合位点，构建了9 个含有不同长度pmel 基因启动子片段的表达载体及1 个核心启动子区突变载体，说明鸡pmel 基因启动子的核心区域为−840–+68 bp，其中−840–−590 bp 和−525–−266 bp 区域为正调控区，−590–−525 bp 区域为负调控区，多态位点 (−456、−435、−410、−374 和−341) 对pmel 基因启动子活性有较大影响。.
Keywords: Bashang long-tail chicken; pmel gene; promoter; transcription factor.