Identification of the core promoter of the pmel gene of Bashang long-tail chickens.

Xiaohui Liu; Rongyan Zhou; Yongdong Peng; Chuansheng Zhang; Lanhui LI; Xianglong LI

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Identification of the core promoter of the pmel gene of Bashang long-tail chickens.

Author: Xiaohui LIU ¹ ; Rongyan ZHOU ¹ ; Yongdong PENG ² ; Chuansheng ZHANG ² ; Lanhui LI ¹ ; Xianglong LI ¹
Author Information

1. College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, Hebei, China.
2. College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066004, Hebei, China.
Publication Type:Journal Article
Keywords: Bashang long-tail chicken; pmel gene; promoter; transcription factor
MeSH: Animals; Chickens; genetics; Cloning, Molecular; CpG Islands; Luciferases; Promoter Regions, Genetic; gp100 Melanoma Antigen; genetics
From: Chinese Journal of Biotechnology 2018;34(11):1750-1759
CountryChina
Language:Chinese
Abstract: 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.