苏淮猪BMP7基因3′-UTR多态性分析  

Polymorphism of the 3′-UTR of BMP7 gene in Suhuai pigs

作　　者：尹杭 杜星[1] 潘增祥[1] 李强 刘红林[1] 李齐发[1] YIN Hang;DU Xing;PAN Zengxiang;LI Qiang;LIU Honglin;LI Qifa(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China;Huaiyin Pig Breeding Farm of Jiangsu Province,Huai’an 223322,China)

机构地区：[1]南京农业大学动物科技学院,江苏南京210095 [2]江苏省淮阴种猪场,江苏淮安223322

出　　处：《南京农业大学学报》2021年第2期346-352,共7页Journal of Nanjing Agricultural University

基　　金：江苏高校“青蓝工程”资助项目(2020)。

摘　　要：[目的]本研究旨在了解苏淮猪骨形态发生蛋白7(BMP7)基因3′-UTR序列特征,以及突变位点多态性与繁殖性状、3′-UTR活性的关系,为解析猪BMP7基因3′-UTR的调控机制提供参考。[方法]采用PCR扩增和测序等技术获得苏淮猪BMP7基因3′-UTR序列,利用生物信息学方法分析其序列特征;池DNA测序筛选苏淮猪BMP7基因3′-UTR上的突变位点,直接测序法对突变位点进行基因分型,采用线性模型分析突变位点多态性与繁殖性状的关系;构建突变位点不同等位型BMP7基因3′-UTR报告载体,用双荧光素酶报告基因系统分析突变位点对BMP7基因3′-UTR活性的影响。[结果]获得639 bp的苏淮猪BMP7基因3′-UTR序列,与其他猪种序列的同源性较高;序列中含有AU-富集元件(ARE)和miRNA应答元件(MRE)等经典的顺式作用元件。在苏淮猪BMP7基因3′-UTR序列的273 nt处发现1个A>G突变,命名为c.*273A>G突变。在苏淮猪群体中BMP7基因c.*273A>G位点有3种基因型,其中AA型为优势基因型,A为优势等位基因。关联分析发现该突变位点多态性与初产产仔数显著关联,其中AG型母猪比GG型每胎高0.83头。荧光素酶活性分析发现A等位型3′-UTR报告载体的荧光素酶活性高于G等位型,但差异不显著。[结论]BMP7是苏淮猪繁殖性状的候选基因。[Objectives]In this study,we aimed to understand the sequence characteristics of the 3′-UTR of BMP7 gene in Suhuai pigs,and the relationship between its polymorphisms and reproductive traits,and 3′-UTR activity,in order to provide reference for investigating the regulatory mechanism of the 3′-UTR of the porcine BMP7 gene.[Methods]The 3′-UTR sequence of Suhuai pig BMP7 gene was obtained by PCR amplification and direct sequencing,and its sequence characteristics were analyzed by bioinformatic methods.Mutations of the 3′-UTR of BMP7 gene in Suhuai pigs were screened by pooled-DNA sequencing,and then genotyped by direct sequencing.Association between polymorphisms and reproductive traits was analyzed by linear model.BMP73′-UTR reporter vectors with different alleles were constructed,and luciferase assays were performed by using a double luciferase reporter gene system.[Results]A 639 bp of 3′-UTR sequence of BMP7 gene was obtained from Suhuai pigs,which had high homology with other pig breeds,and contained classical cis-acting elements such as AU-rich element(ARE)and miRNA response element(MRE).A mutation c.*273A>G was found at 273 nt of the 3′-UTR sequence of BMP7 gene in Suhuai pigs.There were three genotypes in Suhuai pig population,among which AG was the dominant genotype and A was the dominant allele.Association analysis showed that the polymorphisms was significantly associated with the total number of piglets born(TNB)of first parity,in which the sows with genotype AA had 0.83 piglets per litter more than the sows with genotype GG.Furthermore,the luciferase activity of reporter vector of the 3′-UTR with allele A was higher than that of allele G,but the difference was not significant.[Conclusions]BMP7 was a candidate gene for reproductive traits in Suhuai pigs.

关 键 词：苏淮猪 BMP7基因 3′-UTR 多态性 繁殖性状 

分 类 号：S852.23[农业科学—基础兽医学]