绵羊miR-449a/b前体序列组织表达及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2023.06.001

摘要/Abstract

摘要： 【目的】探究miR-449a/b在绵羊生长发育和繁殖性能方面的作用，为进一步研究miR-449a/b在绵羊繁殖调控中的作用提供参考。【方法】以湖羊为研究对象，利用实时荧光定量PCR技术鉴定miR-449a/b在湖羊下丘脑、垂体和卵巢中的相对表达量。使用TargetScan、miRWalk、miRDB软件预测绵羊miR-449a/b前体序列的靶基因，利用微生信和KOBAS网站对其进行GO功能和KEGG通路富集分析。【结果】试验获得的前体序列与NCBI数据库中绵羊前体序列信息比对结果一致。实时荧光定量PCR结果显示，miR-449a在垂体中的相对表达量显著高于miR-449b （P<0.05），miR-449b在卵巢中的相对表达量极显著高于miR-449a （P<0.01）。利用3个靶基因预测软件做韦恩图分析，取其交集得到miR-449a/b的靶基因数目分别为299和23个。GO功能富集显示，miR-449a/b主要被富集到生物进程；KEGG通路富集结果取前20个通路进行分析发现，主要包括催产素、MAPK等与繁殖调控相关的信号通路。【结论】miR-449a在垂体中的表达量较高，miR-449b在卵巢中的表达量较高。miR-449a/b的靶基因数目分别为299和23个，其基因功能主要集中在生物过程，可能参与湖羊的繁殖调控。

关键词: 绵羊; miR-449a; miR-449b; 生物信息学; 靶基因; 繁殖调控

Abstract: 【Objective】 This study was aimed to investigate the role of miR-449a/b in the growth, development and reproductive performance of sheep, and provide reference for further studies on the role of miR-449a/b in reproduction regulation of sheep.【Method】 The relative expression of miR-449a/b in hypothalamus, pituitary and ovary of Hu sheep was studied by Real-time quantitative PCR.TargetScan, miRWalk and miRDB softwares were used to predict the target gene of miR-449a/b precursor sequences in sheep.GO function and KEGG pathway enrichment analysis were performed using bioinformatics and KOBAS websites.【Result】 The comparison result of the precursor sequence obtained from the test was consistent with the precursor sequence information of sheep in NCBI database.Real-time quantitative PCR results showed that the relative expression of miR-449a in pituitary was significantly higher than that of miR-449b (P<0.05), the relative expression of miR-449b in ovary was extremely significantly higher than that of miR-449a (P<0.01).The number of target genes of miR-449a and miR-449b were 299 and 23 by Venn diagram analysis using three target gene prediction softwares, respectively.The results of GO function enrichment showed that miR-449a/b were mainly enriched into biological processes.The results of KEGG pathway enrichment were analyzed by the top 20 pathways, including oxytocin signaling pathway and MAPK signaling pathway and other signaling pathways related to reproductive regulation.【Conclusion】 The expression of miR-449a in pituitary was high, and the expression of miR-449b in ovary was high.The number of target genes of miR-449a/b were 299 and 23, respectively, and its gene function was mainly concentrated in biological process, which might be involved in the reproductive regulation in Hu sheep.

Key words: sheep; miR-449a; miR-449b; bioinformatics; target genes; reproductive regulation

中图分类号:

王赛桥, 赵路, 翟振翰, 张丙雷, 贾万航, 王玉琴. 绵羊miR-449a/b前体序列组织表达及生物信息学分析[J]. 中国畜牧兽医, 2023, 50(6): 2175-2184.

WANG Saiqiao, ZHAO Lu, ZHAI Zhenhan, ZHANG Binglei, JIA Wanhang, WANG Yuqin. Tissue Expression and Bioinformatics Analysis of miR-449a/b Precursor Sequences in Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2175-2184.

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