TGF-β1基因g.8666_8667delAC位点突变与大白猪繁殖性能的关系  

Relationship between mutation g.8666_8667delAC of TGF-β1 gene and reproductive performances in the Large White pig population

作　　者：王玲芳 李尚来 李琦琦 杜星[1] 吴望军[1] 宋奔驰 李齐发[1] WANG Lingfang;LI Shanglai;LI Qiqi;DU Xing;WU Wangjun;SONG Benchi;LI Qifa(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China;Zhengbo Pig Farm of Suyu District,Suqian 223800,China)

机构地区：[1]南京农业大学动物科技学院,江苏南京210095 [2]宿豫区正博生猪养殖场,江苏宿迁223800

出　　处：《南京农业大学学报》2022年第2期351-358,共8页Journal of Nanjing Agricultural University

基　　金：江苏省政策引导类计划(苏北科技专项)(SZ-SQ202044);江苏省农业重大新品种创制专项(PZCZ201734);江苏高校青蓝工程项目(2020)。

摘　　要：[目的]本文旨在了解大白猪转化生长因子β1(TGF-β1)基因第4内含子序列特征,分析其突变位点多态性与繁殖性能的关系及潜在机制。[方法]利用PCR扩增和测序技术获得大白猪TGF-β1基因第4内含子序列,用生物信息学软件分析序列特征;利用DNA混池测序技术筛选目标序列突变位点,用直接测序法进行基因分型,用SAS软件对突变位点多态性与繁殖性能进行关联性分析;利用克隆测序技术测定不同基因型个体卵巢颗粒细胞中TGF-β1基因编码区序列,分析突变对mRNA剪接的影响。[结果]大白猪TGF-β1基因第4内含子序列全长189 bp,含有1个微卫星(GT)_(6)和多个顺式作用元件结合位点,与杜洛克猪相应序列的一致性为98.94%。在第4内含子63~64核苷酸处(TGF-β1基因的8666~8667核苷酸处)发现一个2 bp序列的插入/缺失突变,命名为g.8666_8667delAC。在大白猪群体中发现3种基因型(ins/ins、ins/del和del/del),其中ins/ins为优势基因型(57.8%)。关联性分析发现,ins/del基因型母猪的初产死胎数(NSB)显著高于另2种基因型(P<0.05);del/del基因型母猪的总产仔数(TNB)比ins/ins基因型和ins/del基因型分别高0.47和0.46头(P>0.05)。克隆并测序后发现ins/ins基因型和ins/del基因型母猪卵巢颗粒细胞中TGF-β1基因编码区序列长度一致。[结论]TGF-β1基因是大白猪繁殖性状的候选基因,g.8666_8667delAC位点del/del基因型是总产仔数(TNB)的有利基因型,ins/del基因型母猪初产死胎数(NSB)较高;g.8666_8667delAC位点突变不是通过影响TGF-β1基因的选择性剪接来影响繁殖性能。[Objectives]The paper aimed to understand the sequence characteristics of the intron 4 of TGF-β1 gene in Large White pigs,and analyze the association between the polymorphism and reproductive performance,and the potential mechanism.[Methods]The intron 4 sequence of TGF-β1 gene of Large White pig was obtained by PCR amplification and sequencing,and the sequence characteristics were analyzed using bioinformatics software.Mutations of the target sequence were screened by a pooled DNA sequencing.Direct sequencing was used for genotyping,and SAS software was performed for the association analysis between polymorphism and reproductive performance.The coding sequence of TGF-β1 gene in ovarian granulosa cells(GCs)of sows with different genotypes was determined by clone sequencing,to investigate the effect of mutations on mRNA splicing.[Results]The full-length sequence of the intron 4 of the Large White pig TGF-β1 gene was 189 bp in length,and contained a microsatellite(GT)_(6) and multiple binding sites for cis-acting elements.The intron 4 of the Large White pig TGF-β1 gene shared similarity of 98.94%to Duroc pigs.A 2 bp insertion/deletion mutation was found at nucleotide 63-64 of the intron 4(nucleotide 8666-8667 of TGF-β1 gene),and named g.8666_8667delAC.Three genotypes(ins/ins,ins/del and del/del)were found in the Large White pig population,of which ins/ins genotype was the dominant genotype(57.8%).Association analysis revealed that the number of stillborn(NSB)of sows at first parity with ins/del genotype was significantly higher than the sows carried with ins/ins genotype and del/del genotype.The total number of piglets born(TNB)of sows carried with del/del genotype was 0.47 and 0.46 higher than the ins/ins genotype and the ins/del genotype,respectively.Clone sequencing showed that the length of the coding region of the TGF-β1 gene in GCs of sows carried with the ins/ins genotype and the ins/del genotype were the same.[Conclusions]TGF-β1 gene was a candidate gene for reproductive traits in Large White pigs.Th

关 键 词：大白猪 TGF-Β1基因 多态性 繁殖性能 

分 类 号：S813[农业科学—畜牧学]