乌珠穆沁绵羊RIPK2基因多态性与生长性状的关联  

Association of the RIPK2 Gene Genetic Variation with Ujumqin Sheep Growth Traits

作　　者：马晓萌[1] 轩俊丽[1] 王慧华[1] 袁泽湖[1] 吴明明[1] 朱才业[1] 刘瑞凿 魏彩虹[1] 赵福平[1] 杜立新[1] 张莉[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《中国农业科学》2016年第7期1391-1407,共17页Scientia Agricultura Sinica

基　　金：国家"十二五"863项目(2011AA100307-02);2015年农业部财政项目(2015-ncx-25)

摘　　要：【目的】基于前期绵羊肉用性状GWAS研究结果,旨在探讨RIPK2基因对乌珠穆沁绵羊生长性状的影响,找到该基因中与绵羊生长性状相关的分子标记,同时对GWAS结果进行验证。【方法】在343只乌珠穆沁绵羊试验群体中,选取其中30个个体的血液DNA样本,以相同浓度组成池DNA,设计引物对RIPK2基因外显子及其上下游1 000bp的调控区进行PCR扩增,PCR产物检测为目的条带后测序。使用DNAMAN和Chromas2软件对测序峰图进行分析,采用飞行质谱方法对检测到的SNP位点及前期GWAS得到的SNP位点进行基因型分型。使用Haploview软件对多态位点构建单倍型及连锁不平衡分析。使用SPSS（22.0）软件进行RIPK2基因SNP位点与生长性状间的关联分析。【结果】共发现了4个多态位点,A5536G的同义突变rs01位于第四外显子内,在该位点检测到三种基因型,AA基因型频率为0.42,AG基因型频率为0.45,GG基因型频率为0.13,优势等位基因为A,其频率为0.65;在第七外显子上检测到T8952C错义突变rs02,在该突变位点检测到3种基因型,其中TT基因型频率为0.82,TC基因型频率为0.16,CC基因型频率为0.02,T为优势等位基因,基因频率为0.9;在第十外显子检测到T2836C的突变rs05,在该位点检测到两种基因型TT和CC,没有检测到杂合子基因型,TT基因型频率为0.25,CC基因型频率为0.75;上游调控区发现一个G5181C的突变rs30,有3种基因型,其中GC基因型频率为0.50,CC基因型频率为0.18,GG基因型频率为0.32,G为优势等位基因,其频率为0.58;GWAS得到的SNP位点是T4456C,在本研究中被命名为rs34,在该位点有3种基因型,其中TT基因型频率为0.49,TC基因型频率为0.37,CC基因型频率为0.14,优势等位基因T的频率为0.68。χ^2适合性检验发现,除rs34位点外,其余位点均处于Hardy-Weinberg平衡状态（P〉0.05）。rs02位点处于低度多态（PIC〈0.25）,【Objective】Based on the results of the previous genome-wide association on the sheep meat traits, we studied the possibility of the RIPK2 gene to be a novel candidate gene for the sheep growth traits, in order to confirm the GWAS results, as well as to find new molecular markers related to the sheep growth traits.【Method】In the 343 Ujumqin sheep experiment group, 30 DNA samples with a fixed concentration were selected to construct the DNA pool, primers were designed to extend exons and 1 000 bp of upstream and downstream regulate regions. PCR products were tested and the objective strap were sequenced. The sequence peak maps were analyzed by DNAMAN and Chromas2 softwares, Sequenom Mass-array technology was used to genotype the detected SNP including the SNP obtained by previous GWAS. Haploview was used to construct haplotypes and analyze linkage disequilibrium of the polymorphic loci. Association analysis between the SNPs in RIPK2 gene and the growth traits were performed using SPSS 22.0 software.【Result】 Four SNPs were detected. In exon 4 was a A5536 G synonymous mutation rs01, there were three genotypes on this site and the frequencies of AA, AG and GG were 0.42,0.45 and 0.13 individually, the dominant allele was A with a frequency of 0.65. In exon 7 was a T8952 C missen variation rs02 with three genotypes. The frequencies of TT, TC and CC genotypes were 0.82,0.16 and 0.02. T was the dominant allele and its frequency was 0.9. Rs05 was a T2836 C mutation in exon 10 with only two genotypes TT and CC. The frequency of genotype TT was 0.25 as well as CC was 0.75. In the upstream regions a G5181 C variation rs30 was detected with three genotypes GG, GC and CC. The frequency of heterozygous genotype GC was 0.50 and the other two genotypes CC, GG were 0.18 and 0.32 respectively. G was the dominant allele with its frequency was 0.58.The variation T4456 C detected by GWAS was rs34 in this study and there were three genotypes. The frequency of TT genotype was 0.49, the frequency of TC was 0.37 and the frequen

关 键 词：RIPK2基因 生长性状 多态性 关联分析 乌珠穆沁绵羊 

分 类 号：S826[农业科学—畜牧学]