贵州白水牛ASIP、MC1R和TYR基因多态及其与白毛性状的关联分析
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摘要
ASIP、MC1R和TYR是与动物毛色性状相关的重要候选基因,为了探究ASIP、MC1R、TYR基因多态性及其与贵州白水牛白色被毛之间的关系。本研究以贵州白水牛(20头)和其他毛色水牛(60头)为试验材料,针对3个基因的外显子和部分非编码序列设计引物,利用DNA池作为模板进行扩增测序,对ASIP、MC1R和TYR基因多态性与贵州白水牛被毛之间的关系做了初步探讨。结果发现,在这3个基因中均没有发现特异性的插入/缺失突变,在贵州白水牛ASIP基因的启动子中发现了突变位点:Promoter-A147T和Promoter-A235G;在MC1R基因中筛选到1个错义突变exon1-T843C,TYR基因中筛选出2个错义突变exon1-G826A、exon5-T70C,以及1个同义突变exon3-A83G。这3个基因的SNP位点在扩大样本验证时发现不具有毛色特异性,研究初步表明,ASIP、MC1R、TYR基因的多态性位点不能解释贵州白水牛毛色的特殊变异,为以后对贵州白水牛的毛色的进一步研究奠定了基础。
ASIP, MC1R and TYR are important candidate genes related to animal hair color traits. In order to explore the polymorphism of ASIP, MC1R and TYR genes and the relationship between them and the white coat of Guizhou white buffalo. This research took Guizhou white buffalo(n=20)and other buffaloes(n=60)as experimental materials, designed primers for exons and some non-coding regions of the three genes, then amplified and sequenced them using the DNA pool as template. and preliminarily analyzed the relationship between the polymorphism of ASIP, MC1R and TYR genes and the coat hair of Guizhou white buffalo. The results showed that there are two mutation sites(Promoter-A147 T and Promoter-A235 G) in the promoter of ASIP gene of Guizhou white buffalo. One missense mutation(exon1-T843 C) was detected in MC1R gene, and two missense mutations(exon1-G826 A, exon5-T70 C) and one synonymous mutation(exon3-A83 G) were detected in TYR gene. However, we found that the SNP sites of these three genes were not specific to hair color when the sample size was expanded. This research preliminarily showed that the polymorphism loci of ASIP, MC1R and TYR genes could not explain the special variation of the hair color of Guizhou white buffalo, which laid a foundation for the future research on the hair color of Guizhou white buffalo.
ASIP, MC1R and TYR are important candidate genes related to animal hair color traits. In order to explore the polymorphism of ASIP, MC1R and TYR genes and the relationship between them and the white coat of Guizhou white buffalo. This research took Guizhou white buffalo(n=20)and other buffaloes(n=60)as experimental materials, designed primers for exons and some non-coding regions of the three genes, then amplified and sequenced them using the DNA pool as template. and preliminarily analyzed the relationship between the polymorphism of ASIP, MC1R and TYR genes and the coat hair of Guizhou white buffalo. The results showed that there are two mutation sites(Promoter-A147 T and Promoter-A235 G) in the promoter of ASIP gene of Guizhou white buffalo. One missense mutation(exon1-T843 C) was detected in MC1R gene, and two missense mutations(exon1-G826 A, exon5-T70 C) and one synonymous mutation(exon3-A83 G) were detected in TYR gene. However, we found that the SNP sites of these three genes were not specific to hair color when the sample size was expanded. This research preliminarily showed that the polymorphism loci of ASIP, MC1R and TYR genes could not explain the special variation of the hair color of Guizhou white buffalo, which laid a foundation for the future research on the hair color of Guizhou white buffalo.
引文
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[4] 杨宗禄,梁正文,陶林,等.贵州白水牛品种资源介绍与开发利用[J].畜牧与饲料科学,2014,35(10):33-35.
[5] Maudet C,Taberlet P.Holstein's milk detection in cheeses inferred from melanocortin receptor 1 (MC1R) gene polymorphism[J].J.Dairy Sci.,2002,85(4):707-715.
[6] Hartmeyer M,Scholzen T,Becher E,et al.Human dermal microvascular endothelial cells express the melanocortin receptor type 1 and produce increased levels of IL28 upon stimulation with α-melanocyte stimulating hormone[J].J.Immunol.,1997,159(4):1930-1937.
[7] 刘晓妮,景炅婕,李宝钧,等.ASIP和MC1R基因的研究进展[J].中国草食动物科学,2015,35(1):42-46.
[8] Matsunaga N,Virador V,Santis C,et al.In situ localization of agouti signal protein in murine skin using immunohistochemistry with an ASP-specific antibody[J].Biochem.Biophys.Res.Commun.,2000,270(1):176-182.
[9] Ollmann M M,Barsh G S.Down-regulation of melanocortin receptor signaling mediated by the amino terminus of Agouti protein in Xenopus melanophores[J].J.Biol.Chem.,1999,274(22):15837-15846.
[10] Jackson P J,Douglas N R,Chai B,et al.Structural and molecular evolutionary analysis of agouti and agouti-related proteins[J].Chem.Biol.,2006,13(12):1297-1305.
[11] 徐敏.牛黑素皮质激素受体1(MC1R)基因单核苷酸多态位点(SNP)与毛色表型的分析[D].成都:四川大学,2008.
[12] 敖啟燕,胡玲玲,罗师红,等.不同毛色贵州水牛MC1R、TYR基因SNP研究[J].中国牛业科学,2015,41(4):12-17.
[2] 黄右军,刘业基.广西本地水牛与么拉水牛染色体组型的差异[J].畜牧兽医报,1998,19(4):231-236.
[3] 陈永泽.贵州省畜禽品种志[M].贵阳:贵州科技出版社,1993:20-25.
[4] 杨宗禄,梁正文,陶林,等.贵州白水牛品种资源介绍与开发利用[J].畜牧与饲料科学,2014,35(10):33-35.
[5] Maudet C,Taberlet P.Holstein's milk detection in cheeses inferred from melanocortin receptor 1 (MC1R) gene polymorphism[J].J.Dairy Sci.,2002,85(4):707-715.
[6] Hartmeyer M,Scholzen T,Becher E,et al.Human dermal microvascular endothelial cells express the melanocortin receptor type 1 and produce increased levels of IL28 upon stimulation with α-melanocyte stimulating hormone[J].J.Immunol.,1997,159(4):1930-1937.
[7] 刘晓妮,景炅婕,李宝钧,等.ASIP和MC1R基因的研究进展[J].中国草食动物科学,2015,35(1):42-46.
[8] Matsunaga N,Virador V,Santis C,et al.In situ localization of agouti signal protein in murine skin using immunohistochemistry with an ASP-specific antibody[J].Biochem.Biophys.Res.Commun.,2000,270(1):176-182.
[9] Ollmann M M,Barsh G S.Down-regulation of melanocortin receptor signaling mediated by the amino terminus of Agouti protein in Xenopus melanophores[J].J.Biol.Chem.,1999,274(22):15837-15846.
[10] Jackson P J,Douglas N R,Chai B,et al.Structural and molecular evolutionary analysis of agouti and agouti-related proteins[J].Chem.Biol.,2006,13(12):1297-1305.
[11] 徐敏.牛黑素皮质激素受体1(MC1R)基因单核苷酸多态位点(SNP)与毛色表型的分析[D].成都:四川大学,2008.
[12] 敖啟燕,胡玲玲,罗师红,等.不同毛色贵州水牛MC1R、TYR基因SNP研究[J].中国牛业科学,2015,41(4):12-17.