BRE-AS启动子负调控区变异与启动子活性、母猪繁殖力的关系  

Relationship between the variation of the negative regulatory region ofthe BRE-AS promoter,and promoter activity and sow fertility

作　　者：王杨 王苗苗 单保森 吴望军[1] 杜星[1] 李齐发[1] WANG Yang;WANG Miaomiao;SHAN Baosen;WU Wangjun;DU Xing;LI Qifa(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物科技学院,江苏南京210095

出　　处：《南京农业大学学报》2023年第6期1153-1160,共8页Journal of Nanjing Agricultural University

基　　金：江苏省农业重大新品种创制专项(PZCZ201734);江苏省种业振兴揭榜挂帅项目(JBGS[2021]024);江苏省高校青蓝工程项目(2020)。

摘　　要：[目的]本试验旨在研究BRE反义lncRNA(BRE antisense lncRNA,BRE-AS)基因启动子负调控区(PNRR)序列特征,以及变异位点多态性与母猪繁殖性状、启动子活性的关系。[方法]采用DNA测序法获得大白猪BRE-AS基因PNRR序列;利用池DNA测序筛选大白猪BRE-AS基因PNRR上的变异位点,DNA测序法进行基因分型;利用GLM模型进行变异位点多态性、单倍型与母猪繁殖性状的关联分析;利用荧光素酶报告系统分析变异对BRE-AS基因启动子活性的影响。[结果]大白猪BRE-AS基因的PNRR位于-952~-161 nt,长度792 bp。在大白猪BRE-AS基因PNRR的-794和-756 nt处发现了2个变异位点,分别将其命名为g.-794T>G和g.-756A>C。在大白猪群体中BRE-AS基因g.-794T>G位点有3种基因型,其中TT型为优势基因型(0.752),T为优势等位基因(0.863);在g.-756A>C位点有3种基因型,其中AA型为优势基因型(0.759),A为优势等位基因(0.868);2个变异位点形成3种合并基因型,其中TT/AA型为优势基因型(0.760),T-A为优势单倍型(0.869)。关联分析发现2个位点及其合并基因型均与母猪总产仔数(TNB)、产活仔数(NBA)和健仔数(NSP)等3个繁殖性状间显著(P<0.05)或极显著(P<0.01)关联。荧光素酶活性分析发现G-C单倍型启动子活性极显著高于T-A单倍型(P<0.01)。[结论]BRE-AS是大白猪繁殖性状的候选基因,其PNRR上2个变异位点(g.-794T>G和g.-756A>C)通过影响启动子活性影响其母猪繁殖力。[Objectives]This study was aimed to investigate the sequence characteristics of the promoter negative regulatory region(PNRR)of the porcine BRE antisense lncRNA(BRE-AS)gene,and the relationship between its variant polymorphisms and sow reproductive traits and promoter activity.[Methods]The PNRR sequence of BRE-AS in Yorkshire pigs was obtained by DNA sequencing.Variants in Yorkshire pig BRE-AS PNRR were screened by pooled-DNA sequencing,and genotyped by DNA sequencing.Association analysis of polymorphisms,haplotypes and sow reproductive traits was poformed by using GLM model.The effect of variants on promoter activity of BRE-AS was analyzed by using a luciferase reporter system.[Results]Porcine BRE-AS PNRR was located at-952--161 nt of promoter,with a length of 792 bp.Two variants were found at-794 nt and-756 nt of the BRE-AS PNRR in Yorkshire pigs,which were named as g.-794T>G and g.-756A>C respectively.For SNP g.-794T>G,three genotypes were detected in Yorkshire population,and TT was the dominant genotype(0.752)and T was the dominant allele(0.863).For SNP g.-756A>C,three genotypes were also detected,and AA was the dominant genotype(0.759)and A was the dominant allele(0.868).Two variants formed three combined genotypes,of which TT/AA was the dominant genotype(0.760)and T-A was the dominant haplotype(0.869).Association analysis showed that the two SNP and the combined genotype were significantly(P<0.05 or P<0.01)associated with three reproductive traits,including total number of piglets born(TNB),number of piglets born alive(NBA)and number of healthy piglets(NSP).Double-luciferase activity assays showed that activity of promoter with haplotype G-C was significantly higher than that of promoter with haplotype T-A(P<0.05).[Conclusions]BRE-AS was a candidate gene for sow reproductive traits in Yorkshire pigs,and two variants(g.-794T>G and g.-756A>C)in the PNRR affected sow fertility by affecting the promoter activity.

关 键 词：大白猪 BRE-AS 多态性 繁殖性状 负调控区 

分 类 号：S813[农业科学—畜牧学]