湖羊miR-222前体序列克隆、组织表达及生物信息学分析  

Cloning,Tissue Expression and Bioinformatics Analysis of miR-222 Precursor Sequence in Hu Sheep

作　　者：赵路 王赛桥 张丙雷 贾万航 孙赛祎 杨佳欣 王玉琴[1] ZHAO Lu;WANG Saiqiao;ZHANG Binglei;JIAWanhang;SUN Saiyi;YANG Jiaxin;WANG Yuqin(College of Animal Science and Technology,Henan University of Science and Tecnology,Luoyang 471000,China)

机构地区：[1]河南科技大学动物科技学院,洛阳471000

出　　处：《中国草食动物科学》2024年第4期8-14,共7页China Herbivore Science

基　　金：国家现代农业产业技术体系(CARS-38)。

摘　　要：为探究miR-222在湖羊生长发育和繁殖功能中的作用,试验通过聚合酶链式反应克隆了湖羊miR-222的前体序列;通过实时荧光定量技术检测了miR-222在湖羊下丘脑、垂体和卵巢中的表达规律;使用DNAStar对miR-222成熟序列进行了保守性分析;使用MEGA 11建立了基因进化树,并分析了miRNA-222在各物种间的亲缘关系;通过miRPathD8、TargetScan和StarBase软件预测了湖羊miR-222潜在的靶基因,并对靶基因进行了GO功能富集分析和KEGG通路富集分析。结果表明,试验获得了湖羊miR-222的前体序列;湖羊miR-222在卵巢中的相对表达量显著高于在下丘脑和垂体中的相对表达量(P<0.05);湖羊miR-222的成熟序列与猕猴的亲缘关系最近,在不同物种间具有高度的保守性;湖羊miR-222靶基因预测结果显示,miR-222的潜在靶基因中与繁殖相关的基因有PIK3R1、IGF1、GNAI2和THBS1等;富集到的信号通路包括PI3K-AKT、WnT等,且都在动物繁殖调控中有重要作用。因此,miR-222在湖羊的卵巢发育中可能具有重要的调控作用。To explore the role of miR-222 in the growth,development and reproductive function in this experiment,the precursor sequence of miR-222 in Hu sheep was cloned through polymerase chain reaction.Real-time fluorescent quantitative technology was used to detect the expression pattern of miR-222 in the hypothalamus,pituitary,and ovary of Hu sheep.DNAStar was used to conduct a conservative analysis of the mature sequence of miR-222.MEGA 11 was used to establish a gene evolutionary tree and analyze the phylogenetic relationship of miRNA-222 among various species.miRPathD8,TargetScan,and StarBase software were used to predict the potential target genes of miR-222 in Hu sheep,and GO functional enrichment analysis and KEGG pathway enrichment analysis were conducted on the target genes.The results indicated that the precursor sequence of miR-222 in Hu sheep was successfully obtained.The relative expression level of miR-222 in the ovary of Hu sheep was significantly higher than that in the hypothalamus and pituitary.The mature sequence of miR-222 in Hu sheep was the most closely related to that of macaques,exhibiting a high degree of conservation among different species.Prediction results of the target genes of miR-222 in Hu sheep revealed that some of the potential target genes related to reproduction included PIK3R1,IGF1,GNAI2 and THBS1.The enriched signaling pathways included the PI3K-AKT and Wnt signaling pathways,which both played crucial roles in the regulation of animal reproduction.Therefore,miR-222 may play a crucial regulatory role in the ovarian development of Hu sheep.

关 键 词：湖羊 miRNA 前体序列 生物信息学分析 靶基因 

分 类 号：S826.2[农业科学—畜牧学]