甘南牦牛GDF-10基因多态与生产性状的相关性分析  

GDF-10 Gene Polymorphism and Its Association with Production Traits in Gannan Yak

作　　者：李天科[1,2] 梁春年[1,2] 郎侠[1,2] 裴杰[1,2] 吴晓云[1,2] 褚敏[1,2] 秦文[1,2] 阎萍[1,2] 

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所,兰州730050 [2]甘肃省牦牛繁育工程重点实验室,兰州730050

出　　处：《中国农业科学》2014年第1期161-169,共9页Scientia Agricultura Sinica

基　　金：甘肃牧区生产生态生活优化保障技术集成与示范(2012-2016)(2012BAD13B05);甘肃省科技重大专项计划(1102NKDA027)

摘　　要：【目的】GDF-10基因又称作骨形成蛋白3b,最早通过骨形成蛋白寡聚核苷酸序列的PCR扩增所得,与骨骼的形成和发育有关。很多研究已经表明GDF-10基因在骨骼的形态变化过程中发挥着重要作用。本实验通过研究甘南牦牛GDF-10基因多态性与生产性状间的相关性,证明GDF-10基因与骨骼发育的相关性,发现与牦牛生产性状相关的分子标记,为加快牦牛选育进程,提高牦牛生产性能提供参考。【方法】以298头甘南牦牛血样为材料,随机选取其中的30个DNA样混合组成DNA池,设计9对引物对GDF-10基因外显子1,2和3进行扩增,扩增产物经琼脂糖凝胶回收后测序。运用BLAST和Chromas软件通过测序峰图找出突变位点,然后采用高分辨率熔解曲线分析技术(high resolution melting curve,HRM)进行基因型分型和统计等位基因频率。采用SHEsis和PHASE软件对GDF-10基因多态位点进行配对连锁不平衡和单倍型分析,采用SPSS17.0进行基因多态位点与生产性状关联性分析。【结果】检测到甘南牦牛GDF-10基因外显子3的3个多态位点12116(G/A)、12152(C/T)、和13041(T/C)。群体遗传学分析显示,3个多态位点均表现为低度多态(PIC<0.25);2检验表明牦牛群体在13041(T/C)位点未达到Hardy-Weinberg平衡(P<0.05),其它两个多态位点处于Hardy-Weinberg平衡状态(P>0.05);多态位点配对连锁不平衡分析发现,三个突变位点之间存在弱连锁平衡;关联分析表明,甘南牦牛GDF-10基因不同突变位点的基因型与体斜长、体高、胸围、体重差异显著或极显著(P<0.05或P<0.001),与管围差异不显著(P>0.05);单倍型分析后在群体中发现了7种单倍型组合,其中ATC和ATT组合与牦牛的体尺性状和体重显著相关。【结论】甘南牦牛GDF-10基因3个多态位点和两个单倍型组合与牦牛的体高、体长、体重和胸围显著相关,可以尝试作为牦牛生产性状的候选分子标记,为牦牛遗传资源的保护、开[Objective]Growth differentiation factor-10 （GDF-10） is a member of the TGF-beta super family and the bone morphogenetic protein family （BMP）. It is originally identified by PCR using degenerate oligonucleotides based on the BMP-3 sequence. Studies suggested that the GDF-10 gene plays a key role in skeletal morphogenesis. The objective of this study was to identify the polymorphisms of GDF-10 gene and their correlation with production traits was investigated for the purpose of the breeding of Gannan yak. [Method] Thirty samples were randomly selected from genomic DNA of 298 Gannan yak to construct DNA pool. Nine primers were designed to amplify the exons 1, 2 and 3 of GDF-10 gene. Effective PCR production was purified by DNA gel extraction kit and direct sequencing subsequently. BLAST and Chromas were used to screen the SNPs in GDF-10 gene by the wave height of sequencing map. The genotypes were determined by high-resolution melting curre （HRM） and the frequency of alleles was estimated. The association between genotypes and body measurement was evaluated by software SPSS13.0. Three SNPs （T132C, G672T and G1204A） of GDF-10 gene were identified in the population, which were at extrons 1,extron 2, and extron 3, respectively. The PHASE and SHEsis softwares were used to analyze matching chain disequilibrium and haplotype analysis, respectively. The SPSS17.0 software was used for association analysis.[Result]Three SNPs, 12116 （G/A）, 12152 （C/T） and 13041 （T/C） were discovered in exon 3. Through population genetics analysis, the results showed that three locals were at low polymorphic status （PIC＆lt;0.25）. Theχ2 tests showed that the locus of 13041 （T/C） was not all in the status of Hardy-Weinberg equilibrium （P＆lt;0.05）, the other two loci were all in the status of Hardy-Weinberg equilibrium （P＆gt;0.05）. Analysis of matching chain disequilibrium and haplotype analysis show that there is a weak chain of balance between the three locals. Analysis of association of polym

关 键 词：牦牛 GDF-10基因 多态性 高分辨率溶解曲线 单倍型 生产性状 

分 类 号：S823.85[农业科学—畜牧学]