绵羊INSL3基因组织表达规律与潜在功能位点分析研究  

Tissue Expression Pattern and Functional SNP Analysis of INSL3 Gene in Sheep

作　　者：潘章源 张自杰[1] 季久秀 李华振 孙安芳 李富宽 王慧 储明星 吕慎金[1] PAN Zhangyuan;ZHANG Zijie;JI Jiuxiu;LI Huazhen;SUN Anfang;LI Fukuan;WANG Hui;CHU Mingxing;L Shenjin(College of Agriculture and Forestry Science,Linyi University,Linyi 276005,China;Key Laboratory of Animal Genetics,Breeding and Reproduction of Ministry of Agriculture and RuralAffairs Institute of Animal Science Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]临沂大学农林科学学院,临沂276005 [2]中国农业科学院北京畜牧兽医研究所,农业农村部动物遗传育种与繁殖重点实验室,北京100193

出　　处：《畜牧兽医学报》2020年第4期679-687,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31802031;31872398);山东省自然科学基金(ZR2018BC045);地方高校国家级大学生创新创业训练计划项目(201710452041);临沂大学博士启动基金(LYDX2016BS068);动物行为福利与健康养殖技术技能型创新团队。

摘　　要：旨在对绵羊INSL3基因组织表达规律与潜在功能位点进行分析。本研究首先利用RT-PCR和实时荧光定量PCR技术对健康、角型正常、1岁的小尾寒羊成年母羊和公羊各3只的INSL3组织表达规律进行研究,随后利用10个品种重测序数据分析INSL3基因区域主成分(PCA)和功能位点。定量结果显示,INSL3基因在卵巢和睾丸中表达最高,在软角也表达,公羊睾丸表达量极显著高于母羊卵巢组织(P<0.01),公羊软角组织表达显著高于母羊(P<0.05),结合公母羊角的大小差异,说明该基因可能与角有关。PCA结果显示,该区域一定程度上按角的有无进行品种聚类,进一步说明该区域可能与角有关。功能位点分析发现,4个潜在SNPs位点在有角和无角群体中存在显著性差异分布,其中SNP1处于KDM1A转录因子结合位点上,SNP3位于ESR1结合位点,SNP6存在于终止子区域。同时还发现一个错义突变和同义突变。本研究表明,绵羊INSL3基因可能既与公母羊角的差异有关,也可能与绵羊角的有无有关,并找到多个潜在功能位点,为今后研究其功能机制奠定基础。The aim of this study was to analyze the tissue expression pattern and functional SNP of INSL3 gene in sheep.In this study,three each of male and female 1-year-old Small Tail Han sheep with healthy body condition and normal horn were used to detect tissue expression pattern of INSL3 by RT-PCR and real-time PCR.The gene resequencing data of 10 sheep breeds were used to explore the principal component analysis(PCA)and SNP of INSL3 gene region.The qPCR result showed that the expression level of INSL3 gene was the highest in ovary and testis,and INSL3 gene was also expressed in soft horn.The expression of INSL3 gene in testis was significantly higher than that in ovary(P<0.01),and the expression of INSL3 gene in soft horn of ram was significantly higher than that in ewe(P<0.05).PCA results showed that the INSL3 gene region was clustered according to the presence or absence of horn in a certain extent,which further indicated that this region might be related to horn.Functional site analysis revealed that 4 potential SNP sites were significantly different in distribution in horned and hornless populations.SNP1 was at the transcription factor binding site of KDM1A,SNP3 at the ESR1 binding site,and SNP6 at the terminator region.A missense mutation and a synonymous mutation were also found.This study shows that INSL3 gene may be related to the different horn size between male and female sheep as well as presence or absence of the horn,and multiple potential functional sites have been found.This study provides the basis for the future research on the functional mechanism of INSL3.

关 键 词：INSL3 实时荧光定量PCR 组织表达谱 功能位点 绵羊 

分 类 号：S826.2[农业科学—畜牧学]