绵羊TrkA基因的表达及其SNPs与产羔性状的关联性分析

发布时间：2019-01-10 16:29

【摘要】：为研究绵羊(Ovis aries)酪氨酸蛋白激酶受体(tyrosine kinase A,Trk A)基因组织表达特征及其SNPs与产羔性状的关联性,本研究以Trk A基因为研究对象,利用q RT-PCR分析其在湖羊心、肝、脾、肺、肾、瘤胃、十二指肠、背最长肌、脂肪、下丘脑、垂体和卵巢等12种组织中的表达水平,同时采用DNA混合池测序和限制性长度片段多态性(restriction fragment length polymorphism,RFLP)等技术检测Trk A基因在963只有产羔记录湖羊(n=526)和小尾寒羊(n=437)群体中的多态性,及其与产羔性状的关联性。结果表明,Trk A基因在卵巢、肺和肝脏中的表达水平最高,在心脏中次之,在背最长肌、瘤胃、脂肪、下丘脑、垂体、脾、肾和十二指肠间表达量较低。通过DNA池测序和RFLP分析,在Trk A基因的内含子9和14分别检测到NC_019458.2:g.105281586CT和NC_019458.2:g.105284246GC两个突变。在NC_019458.2:g.105281586CT位点,小尾寒羊和湖羊群体中均检测到3种基因型,分别为CC、CT和TT型,其中CT为优势基因型,C为优势等位基因;在该位点,湖羊群体遗传杂合度(heterozygosity,He)和遗传纯合度(homozygosity,Ho)均为0.50,有效等位基因数(effective allele numbers,Ne)为1.99;小尾寒羊He=0.51,Ho=0.49,Ne=1.95;多态信息含量(polymorphic information content,PIC)均为0.37,且在小尾寒羊群体中偏离了Hardy-Weinberg平衡状态。在NC_019458.2:g.105284246GC位点上同样检测到GG、GC和CC三种基因型,其中GC为优势基因型,C为优势等位基因;该位点在湖羊群体He和Ho均为0.50,Ne=2.00;小尾寒羊He=0.49,Ho=0.51,Ne=1.96。PIC均为0.37;χ2适合性检验结果表明,湖羊群体在该位点显著偏离于HardyWeinberg平衡状态。多态位点与绵羊产羔数的关联性分析表明,NC_019458.2:g.105281586CT与湖羊和小尾寒羊的产羔数显著相关(P0.05),而NC_019458.2:g.105284246GC突变位点仅与湖羊的产羔性能显著相关(P0.05)。本研究结果表明,Trk A基因可作为绵羊产羔性状早期筛选的候选基因,其SNPs可为绵羊繁殖性状分子标记辅助选择提供依据。
[Abstract]:In order to study the expression characteristics of sheep (Ovis aries) tyrosine protein kinase receptor (tyrosine kinase (tyrosine kinase Trk A gene and the relationship between SNPs and lambing traits, the Q RT-PCR was used to analyze the expression of Trk A gene in the heart and liver of Hu sheep. The expression levels of spleen, lung, kidney, rumen, duodenum, longissimus dorsi muscle, fat, hypothalamus, pituitary and ovary were detected by DNA mixed pool sequencing and restriction fragment polymorphism (restriction fragment length polymorphism,). RFLP and other techniques were used to detect the polymorphism of Trk A gene in 963 lambing only recorded populations of Hu Sheep (nong526) and small tail Han Sheep (nb437), and their association with lambing traits. The results showed that the expression of, Trk A gene was the highest in ovary, lung and liver, followed by heart, and lower in longissimus dorsi, rumen, fat, hypothalamus, pituitary, spleen, kidney and duodenum. NC_019458.2:g.105281586CT and NC_019458.2:g.105284246GC mutations were detected in intron 9 and 14 of Trk A gene by DNA pool sequencing and RFLP analysis, respectively. At the NC_019458.2:g.105281586CT locus, three genotypes, CC,CT and TT, were detected in small tail Han sheep and Hu sheep, respectively, in which CT was the dominant genotype and C was the dominant allele. At this locus, the genetic heterozygosity (heterozygosity,He) and genetic homozygosity (homozygosity,Ho) of Hu sheep population were 0.50, and the effective allele number (effective allele numbers,Ne was 1.99.The small tail Han sheep was He=0.51,Ho=0.49,Ne=1.95;. The polymorphic information content (polymorphic information content,PIC) was 0.37, and deviated from the Hardy-Weinberg equilibrium state in the small tail Han sheep population. Three genotypes of GG,GC and CC were also detected at the NC_019458.2:g.105284246GC locus, in which GC was the dominant genotype and C was the dominant allele, and this locus was 0.50 Nefeng 2.00 in both He and Ho of Hu sheep population. The results of 蠂 2 fitness test showed that the population of Hu sheep deviated significantly from the equilibrium state of HardyWeinberg at this site. The correlation analysis between polymorphic loci and lambing size showed that NC_019458.2:g.105281586CT was significantly correlated with litter size of Hu sheep and small tail Han sheep (P0.05). However, the NC_019458.2:g.105284246GC mutation site was only significantly correlated with the lambing performance of Hu sheep (P0.05). The results showed that, Trk A gene could be used as a candidate gene for early screening of lamb traits and its SNPs could provide a basis for molecular marker-assisted selection of sheep reproductive traits.
【作者单位】：甘肃农业大学动物科学技术学院;青海大学农牧学院;兰州大学草地农业科技学院;甘肃省肉羊繁育生物技术工程实验室;
【基金】：国家自然科学基金项目(No.31472072) 国家肉羊现代肉羊产业技术体系建设专项(No.CARS-39)
【分类号】：S826

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