miR-1-3p对乾华肉用美利奴羊次级毛囊发育相关基因的调控研究  

Regulation of miR-1-3p on Genes Related to Secondary Hair Follicle Development in Qianhua Mutton Merino Sheep

作　　者：张桂山 张英楠 杨树宝 徐晶 姜怀志[3] ZHANG Guishan;ZHANG Yingnan;YANG Shubao;XU Jing;JIANG Huaizhi(College of Public Health,Jilin Medical University,Jilin,Jilin 132013,China;Basic Medical College,Jilin Medical University,Jilin,Jilin 132013,China;College of Animal Science and Technology,Jilin Agricultural University,Changchun,Jilin 130118,China)

机构地区：[1]吉林医药学院公共卫生学院,吉林吉林132013 [2]吉林医药学院基础医学院,吉林吉林132013 [3]吉林农业大学动物科学技术学院,吉林长春130118

出　　处：《家畜生态学报》2023年第10期9-13,共5页Journal of Domestic Animal Ecology

基　　金：吉林省教育厅科学技术研究项目(JJKH20220471KJ);吉林医药学院博士科研启动基金项目(JYBS2021015LK,JYBS2021017LK);吉林省科技发展计划项目(YDZJ202201ZYTS220)。

摘　　要：为了探讨miR-1-3p及其靶基因与乾华肉用美利奴羊次级毛囊发生发育关系,该研究采集2周岁乾华肉用美利奴羊皮肤样品,利用生物信息学方法预测miR-1-3p的靶基因,荧光定量PCR检测miR-1-3p及其靶基因相对表达量,通过Western blot对miR-1-3p的靶基因成纤维细胞生长因子14(fibroblast growth factor 14,FGF 14)进行蛋白定量检测,并利用双荧光素酶报告基因试验验证miR-1-3p与候选靶基因的靶向关系。结果表明:通过Targetscan v7.0、miRanda和RNA223个靶基因预测软件确定FGF 14为miR-1-3p的候选靶基因;miR-1-3p在毛囊生长期低表达,在休止期高表达;FGF 14基因mRNA在毛囊生长期高表达,休止期低表达,与FGF 14蛋白表达规律一致。双荧光素酶报告基因试验结果表明miR-1-3p的靶基因为FGF 14。可见,乾华肉用美利奴羊次级毛囊miR-1-3p与FGF 14存在靶向调控关系,miR-1-3p可能是通过负调控FGF 14实现对乾华肉用美利奴羊次级毛囊发生发育进行调控。The objective of this experiment was to study the relationship between miR-1-3p and its target genes and secondary hair follicles development in Qianhua mutton merino sheep.Bioinformatics was used to predict the target genes of miR-1-3p;fluorescence quantitative PCR was adopted to detect the relative expression levels of miR-1-3p and its target genes;Western blot was used to determine the protein quantification of Fibroblast Growth Factor 14(FGF_(14))protein of miR-1-3p target gene;Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-1-3p and candidate target genes.FGF 14 was identified as the candidate target gene of miR-1-3p by Targetscan v7.0,miRanda and RNA22 software.MiR-1-3p was of low expression in anagen and high expression in telogen.On the contrary,the relative expression of FGF_(14) mRNA was high in anagen and low in telogen,which was consistent with the expression pattern of FGF_(14) protein.The results of the double luciferase reporter gene assay showed that the target gene of miR-1-3p was FGF_(14).In conlusion,there is a targeted regulatory relationship between miR-1-3p and FGF 14 in secondary hair follicles of Qianhua mutton merino sheep,and miR-1-3p may regulate secondary hair follicles development in Qianhua mutton merino sheep by negatively regulating FGF_(14).

关 键 词：乾华肉用美利奴羊 次级毛囊 miR-1-3p FGF 14 

分 类 号：S826.2[农业科学—畜牧学]