Liu Xiaohui, Zhou Rongyan, Peng Yongdong, Zhang Chuansheng, Li Lanhui, Li Xianglong
College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, Hebei, China.
College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066004, Hebei, China.
Sheng Wu Gong Cheng Xue Bao. 2018 Nov 25;34(11):1750-1759. doi: 10.13345/j.cjb.180050.
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.
为探究坝上长尾鸡pmel核心启动子的活性，我们构建了双荧光素酶表达载体，并用Lipofectamine 2000将其瞬时转染至DF1细胞中。我们使用双荧光素酶检测试剂盒测量荧光素酶活性。克隆了坝上长尾鸡pmel基因5′侧翼区的1268 bp片段。从 -1200 bp至 +68 bp的区域包含2个CpG岛和多个转录因子结合位点。我们构建了9个具有不同启动子区域的表达载体以及坝上长尾鸡pmel基因核心启动子区域的突变载体。在pmel基因中鉴定出了从 -840 bp至 +68 bp的核心启动子区域。从 -590至 -525 bp的区域在转录过程中对pmel基因起负调控作用。-840--590 bp和 -525--266 bp区域为正调控区域。多态性位点（-456、-435、-410、-374和 -341）对pmel基因的启动子活性有显著影响。
