牦牛Ihh基因组织表达分析、SNP检测及其基因型组合与生产性状的关联分析

doi:10.11843/j.issn.0366-6964.2015.01.007

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (1): 50-59.doi: 10.11843/j.issn.0366-6964.2015.01.007

牦牛Ihh基因组织表达分析、SNP检测及其基因型组合与生产性状的关联分析

李天科^1,2,3，赵娟花^1,2，裴杰^1,2，梁春年^1,2，郭宪^1,2，秦文^1,2,3，阎萍^1,2,3*

（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； 3.甘肃农业大学动物科学技术学院，兰州 730070）

收稿日期:2014-01-25 出版日期:2015-01-23 发布日期:2015-01-23
通讯作者: 阎萍，研究员，博士生导师，主要从事动物遗传育种与繁殖研究，E-mail:pingyan@sohu.com
作者简介:李天科（1987-），男，硕士生，主要从事动物遗传育种与繁殖研究，E-mail:litianke1987@163.com
基金资助:
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）

Tissue Expression，SNP Detection and Association of Genotype Combination of Ihh Gene with Production Traits in Yak

LI Tian-ke^1,2,3，ZHAO Juan-hua^1,2，PEI Jie^1,2，LIANG Chun-nian^1,2，GUO Xian^1,2，QIN Wen^1,2,3，YAN Ping^1,2,3*

(1.Lanzhou Institute of Husbandry and Pharmaceutical Sciences，Chinese Academy of Agricultural Sciences，Lanzhou 730050，China;2.Key Laboratory of Yak Breeding Engineering，Lanzhou 730050，China;3.College of Animal Science and Technology，Gansu Agricultural University，Lanzhou 730070，China)

Received:2014-01-25 Online:2015-01-23 Published:2015-01-23

摘要/Abstract

摘要：

为了研究牦牛Ihh基因多态位点基因型组合与生产性状间的相关性，发现与牦牛生产性状相关的分子标记，同时进一步研究Ihh基因在牦牛和黄牛各个组织的分布和表达，试验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，同时，运用荧光定量PCR 分析Ihh基因在牦牛和黄牛不同器官的表达差异。试验结果表明，Ihh基因在牦牛和黄牛的所有组织中均表达。然后采用SHEsis和PHASE软件对Ihh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进行多态位点单倍型组合与生产性状关联性分析。结果检测到牦牛Ihh基因外显子3的 2个多态位点5855（C/T）和6383（G/A）。群体遗传学分析显示，2个多态位点均表现为高度多态（PIC>0.25）；χ²检验表明，甘南牦牛和大通牦牛群体在两个突变位点处于Hardy-Weinberg平衡状态（P＞0.05），而天祝牦牛在2个突变位点处未达到Hardy-Weinberg平衡（P＜0.05）；多态位点配对连锁不平衡分析发现，2个突变位点之间存在强连锁平衡，有4种单倍型组合；基因型组合主要发现了3种。关联分析表明，Ihh基因2个突变位点的3种基因型组合对牦牛体斜长、体高、胸围、管围和体重有显著差异（P＜0.05），具有CTGA基因型组合的牦牛个体在体斜长、体高、胸围、管围和体重方面显著高于CCGG和TTAA型（P＜0.05）。因此，综上推断Ihh基因基因型组合与牦牛体斜长、体高、胸围、管围和体重存在相关性。

Abstract:

The objective of this study was to identify the polymorphisms of Ihh gene and their correlation with production traits，then to detect the molecular markers related to production traits，and investigate distribution and expression in tissues detected in yak and cattle.At the same time，RT-PCR method was used to detect the expression of Ihh gene in different tissues in yak and cattle.BLAST and Chromas were used to screen the SNPs in Ihh gene by the wave height of sequencing map.The genotypes were determined by high-resolution melting curve(HRM) and the alleles frequency was estimated.The PHASE and SHEsis softwares were used to analyze matching chain disequilibrium and haplotype，respectively.The SPSS17.0 software was used for association analysis.Two SNPs(5855(C/T) and 6383(G/A)) of Ihh gene were indentified in the population，which were on exon 3.Through population genetics analysis，the results showed that 2 loci were at high polymorphic status(PIC>0.25).The χ²tests showed that the 2 loci were in the status of Hardy-Weinberg equilibrium(P＜0.05) in Gannan and Datong yaks; the 2 loci were not all in the status of Hardy-Weinberg equilibrium(P＞0.05) in Tianzhu yak.Analysis of matching chain disequilibrium and haplotype showed that there was a strengh linkage equilibrium between the 2 loci with 4 haplotype combinations and 3 genotype combinations.Analysis of association of polymorphism with body measurement traits at all mutation loci showed that 3 genotype combinations had significant effects on body height，body length，weight，tube girth and herrt girth(P＜0.05).Yak with CTGA genotype combination had higher body height，body length，weight，tube girth and herrt girth than individuals with CCGG and TTAA.The result of expression showed that Ihh gene was expressed in 8 different tissues.The results suggest that Ihh gene may have potential effects on production traits in the above mentioned yak populations and could be used for marker-assisted selection.

中图分类号:

李天科，赵娟花，裴杰，梁春年，郭宪，秦文，阎萍. 牦牛Ihh基因组织表达分析、SNP检测及其基因型组合与生产性状的关联分析[J]. 畜牧兽医学报, 2015, 46(1): 50-59.

LI Tian-ke，ZHAO Juan-hua，PEI Jie，LIANG Chun-nian，GUO Xian，QIN Wen，YAN Ping. Tissue Expression，SNP Detection and Association of Genotype Combination of Ihh Gene with Production Traits in Yak[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(1): 50-59.

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牦牛Ihh基因组织表达分析、SNP检测及其基因型组合与生产性状的关联分析

Tissue Expression，SNP Detection and Association of Genotype Combination of Ihh Gene with Production Traits in Yak

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