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

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.