中国动物传染病学报2024，Vol.32Issue(3)：154-163,10.
山羊NPC1基因核心启动子鉴定与转录调控分析
Identification and Analysis of the Core Region of Goat NPC1 Gene Promoter
摘要
Abstract
Niemann-pick C1(NPC1)encoded by the NPC1 gene participates in lipid raft formation,endocytosis of pathogenic microorganisms and intracellular transport of endosomes.We aimed to study the transcriptional regulation mechanism of NPC1 gene in Hainan black goats and to provide a theoretical reference for studying the interactions between pathogenic microorganisms and host cells.First,the 1466 bp promoter sequence of the goat NPC1 gene was analyzed and cloned for the study.The 5'-end truncated NPC1 promoters were amplified and constructed into the dual-luciferase reporter vector to screen the core promoter region of the NPC1 gene.Then,the key transcription factors in the core promoter region were further predicted and analyzed and the deletion vectors of transcription factor binding sites were constructed to screen the key transcription factor binding sites of NPC1 gene.The results showed that the core region of goat NPC1 gene promoter located from-195 bp to-60 bp upstream of the transcription initiation site and there were E2F3(-134/-120 bp),SREBP-1(-107/-96 bp)SP1(-68/-62 bp)and other transcription factor binding sites.The results of the dual-luciferase reporter assay showed that deletion of three transcription factor binding sites E2F3,SREBP-1 and SP1 significantly decreased the transcriptional activity of goat NPC1 gene promoter(P＜0.01)and deletion of SREBP-1 binding site significantly decreased the transcriptional activity of NPC1 gene.These results suggested that SREBP-1 transcription factor played an important role in the transcriptional activity of NPC1 gene in Hainan black goats.In this experiment,identification of the core region(-195/-60 bp)of goat NPC1 gene promoter and the key transcription factor binding site SREBP-1 in this region gained better understanding of the molecular mechanism of goat NPC1 gene-mediated interaction between pathogenic microorganisms and hosts.关键词
海南黑山羊/NPC1基因/转录调控/核心启动子/SREBP-1Key words
Hainan black goat/NPC1 gene/transcriptional regulation/core promoter/SREBP-1分类
农业科技引用本文复制引用
李崇瑞,王凤阳,陈思,翟哲,王雪梅,吴艳茹,刘志勇,蒋俊明,满初日嘎,杜丽..山羊NPC1基因核心启动子鉴定与转录调控分析[J].中国动物传染病学报,2024,32(3):154-163,10.基金项目
国家自然科学基金项目(32160831) （32160831）
国家肉羊产业技术体系(财政部和农业农村部CARS38) （财政部和农业农村部CARS38）
海南省院士创新平台科研专项(YSPTZX202013) （YSPTZX202013）
海南大学科研启动基金项目(KYQD(ZR)1946) （KYQD(ZR）
