角蛋白关联蛋白基因（KAP和KRT）与绵羊毛品质的相关性研究
摘要
新疆是我国重要的绵羊生产基地,细毛羊数量居全国首位,细毛羊业在国内处于领先地位,目前国内外毛纺产品也已经向轻薄、柔软、挺括、高档方向发展。因此,加快细毛羊的发展,提高细毛羊的育种水平,既是社会经济发展的需要,也是此领域科技进步的需要。但从目前情况来看,要提高或保持现有高品质细毛羊的羊毛品质存在一定难度,其关键问题是对控制绵羊主要经济性状的基因调控机制研究较少,特别是与绵羊毛品质相关的基因位点及相关关系尚不清楚。目前研究热点主要是针对构成羊毛纤维的角蛋白关联蛋白(Keratin-associated proteins,KAPs)和角蛋白中间丝(Keratin intermediate filament, KRT-IF)的相关基因的研究上,本研究在已有研究的基础上进一步从两个方面对KAP和KRT相关基因进行了研究:
一、新疆5种绵羊角蛋白关联蛋白基因遗传多态性研究
本试验选择KAP和KRT部分基因(KAP1.3,KAP6-1,KAP3.2,KAP8和KRT1.2)作为候选基因,利用primer5.0引物设计软件对相关基因分别设计13对引物,以新疆不同毛品质的绵羊(半细毛陶赛特羊;半细毛羊萨福克羊;肉羊德克塞尔羊;粗毛羊湖羊;中国美利奴(新疆军垦型)细毛羊)为研究基础,利用PCR-RFLP和PCR-SSCP技术进行遗传多态性分析。结果得出:(1)5种绵羊在KAP1.3位点都处于中度多态PIC在0.25～0.50之间;χ2适合性检验德克塞尔羊未达到遗传平衡,其他四种绵羊均达到平衡。(2)在KAP81位点陶赛特羊为高度多态(PIC>0.50),德克塞尔羊处于低度多态(PIC<0.25),中国美利奴羊,萨福克羊和湖羊处于中度多态;经χ2适合性检验得出,只有德克塞尔羊达到遗传平衡,其他四种绵羊均未达到平衡。(3)在KAP87位点5种绵羊的多态信息含量较低,萨福克羊为中度多态,其他4种绵羊均为低度多态;χ2适合性检验显示萨福克羊未达到遗传平衡,其他4种绵羊达到遗传平衡。(4)在KRT123位点5种绵羊中湖羊为中度多态,其他4种绵羊均为低度多态;χ2适合性检验显示5种绵羊达到遗传平衡。(5)X611位点5种绵羊中德克塞尔羊处于中度多态,其他4种绵羊均处于高度多态;χ2适合性检验5种绵羊均未达到遗传平衡。因此KAP1.3,KAP81,KAP87,KRT123和X611位点可能与毛品质存在相关性。
二、毛细度,强度,长度育种值与基因位点相关性研究
利用第一部分分析得出与绵羊毛品质相关的基因位点,以中国美利奴(新疆军垦型)细毛羊为研究基础,利用多重比较及t检验计算分析个体毛性状育种值与基因位点的关系得出:(1)表型值和个体育种值分析结果:KAP81位点的AC基因型,KAP1.3位点的AA基因型和X611位点的BC基因型可对羊毛细度进行标记;KAP81位点的BC基因型和KRT123位点的AB基因型可对羊毛强度进行标记;X611位点的AA基因型可对羊毛长度进行标记。(2)对15只公羊利用分子标记检测结果进行育种值分析结果:通过基因型效应估计育种值,其结果与后代表型值更接近,因此可以直接利用基因型进行优质绵羊的选育,并可通过同型交配达到快速选种的目的。
Xinjiang, an important base of sheep breeding, owing the most, is the important product-place of fine-wool and in the leading place in China.At present,domestic and international wool textile products have also to the light,soft,straight out,high-grade direction.So it is necessary to raise fine wool sheep breeding, not only for society economic but also for technology ascensive in breeding.But now, it is difficult to raise or maintain the quality of present superior fine-wool, since it is studied less about the mechanism of gene regulation, controlling the main economy traits, especially, it is dimness about the gene that have correlation with wool trait of sheep ,and we know little about they correlativity. At present the research is focus on the gene that relation with Keratin-associated proteins and Keratin intermediate filament, and the protein was construct the hair fiber. We study KAP and KRT associate gene in two way on the base of already research.
Firstly, Study the genetic polymorphism of KAP and KRT gene and 5 sheep in Xinjiang
Choosing KAP and KRT gene (KAP1.3,KAP6-1,KAP3.2,KAP8 and KRT1.2) as candidate gene, the genetic relationship between KAP and KRT gene and different wool trait of sheep (Chinese Merino sheep, Dorset sheep, Suffolk sheep, Texel, Hu sheep) in Xinjiang were analyzed by PCR-SSCP and PCR-RFLP method. The result showed:(1) 5 different sheep was midrange polymorphism at KAP1.3 loci, and PIC was between 0.25 and 0.5. Texel sheep not achieve genetic balance tested byχ2. (2) KAP81 loci Dorset sheep was high polymorphism (PIC>0.5), Texel sheep was low polymorphism (PIC<0.25), Chinese Merino sheep, Hu sheep and Suffolk sheep was midrange polymorphism (0.25genetic balance tested byχ2,other 4 sheep were not achieve genetic balance.(3) KAP87 loci Suffolk sheep was midrange polymorphism, other 4 sheep was low polymorphism. tested byχ2 the 5 sheep had achieve genetic balance.(4) KRT123 loci Hu sheep was midrange polymorphism, and other 4 sheep was low polymorphism, tested byχ2 the 5 sheep had achieve genetic balance.(5) X611 loci Texel was midrange polymorphism, other 4 sheep was high polymorphism.5 sheep weren’t achieve genetic balance tested byχ2.So KAP1.3,KAP6-1,KAP3.2,KAP8 and KRT1.2 may have relation with wool trait.
Secondly, study the relation of gene and wool fineness, strength and length breeding value
Study the relationship between Chinese Merino sheep wool traits breeding value and gene loci by multiple comparison and t tset, gene has relation of wool trait in different sheep, research in first part. The result showed that:(1)result of phenotypic value and individual breeding value: AC genotype of KAP81,AA genotype of KAP1.3,BC genotype of X611,could mark wool fineness; BC genotype of KAP81, AB genotype of KRT123 could mark wool strength; AA genotype of X611 could mark wool length. (2) The result of Study the genotype breeding value of 15 Sire sheep was: It is closer between genotype breeding value and phenotypic value of progeny .So it is convenience use homogamy to get high quality sheep.
一、新疆5种绵羊角蛋白关联蛋白基因遗传多态性研究
本试验选择KAP和KRT部分基因(KAP1.3,KAP6-1,KAP3.2,KAP8和KRT1.2)作为候选基因,利用primer5.0引物设计软件对相关基因分别设计13对引物,以新疆不同毛品质的绵羊(半细毛陶赛特羊;半细毛羊萨福克羊;肉羊德克塞尔羊;粗毛羊湖羊;中国美利奴(新疆军垦型)细毛羊)为研究基础,利用PCR-RFLP和PCR-SSCP技术进行遗传多态性分析。结果得出:(1)5种绵羊在KAP1.3位点都处于中度多态PIC在0.25～0.50之间;χ2适合性检验德克塞尔羊未达到遗传平衡,其他四种绵羊均达到平衡。(2)在KAP81位点陶赛特羊为高度多态(PIC>0.50),德克塞尔羊处于低度多态(PIC<0.25),中国美利奴羊,萨福克羊和湖羊处于中度多态;经χ2适合性检验得出,只有德克塞尔羊达到遗传平衡,其他四种绵羊均未达到平衡。(3)在KAP87位点5种绵羊的多态信息含量较低,萨福克羊为中度多态,其他4种绵羊均为低度多态;χ2适合性检验显示萨福克羊未达到遗传平衡,其他4种绵羊达到遗传平衡。(4)在KRT123位点5种绵羊中湖羊为中度多态,其他4种绵羊均为低度多态;χ2适合性检验显示5种绵羊达到遗传平衡。(5)X611位点5种绵羊中德克塞尔羊处于中度多态,其他4种绵羊均处于高度多态;χ2适合性检验5种绵羊均未达到遗传平衡。因此KAP1.3,KAP81,KAP87,KRT123和X611位点可能与毛品质存在相关性。
二、毛细度,强度,长度育种值与基因位点相关性研究
利用第一部分分析得出与绵羊毛品质相关的基因位点,以中国美利奴(新疆军垦型)细毛羊为研究基础,利用多重比较及t检验计算分析个体毛性状育种值与基因位点的关系得出:(1)表型值和个体育种值分析结果:KAP81位点的AC基因型,KAP1.3位点的AA基因型和X611位点的BC基因型可对羊毛细度进行标记;KAP81位点的BC基因型和KRT123位点的AB基因型可对羊毛强度进行标记;X611位点的AA基因型可对羊毛长度进行标记。(2)对15只公羊利用分子标记检测结果进行育种值分析结果:通过基因型效应估计育种值,其结果与后代表型值更接近,因此可以直接利用基因型进行优质绵羊的选育,并可通过同型交配达到快速选种的目的。
Xinjiang, an important base of sheep breeding, owing the most, is the important product-place of fine-wool and in the leading place in China.At present,domestic and international wool textile products have also to the light,soft,straight out,high-grade direction.So it is necessary to raise fine wool sheep breeding, not only for society economic but also for technology ascensive in breeding.But now, it is difficult to raise or maintain the quality of present superior fine-wool, since it is studied less about the mechanism of gene regulation, controlling the main economy traits, especially, it is dimness about the gene that have correlation with wool trait of sheep ,and we know little about they correlativity. At present the research is focus on the gene that relation with Keratin-associated proteins and Keratin intermediate filament, and the protein was construct the hair fiber. We study KAP and KRT associate gene in two way on the base of already research.
Firstly, Study the genetic polymorphism of KAP and KRT gene and 5 sheep in Xinjiang
Choosing KAP and KRT gene (KAP1.3,KAP6-1,KAP3.2,KAP8 and KRT1.2) as candidate gene, the genetic relationship between KAP and KRT gene and different wool trait of sheep (Chinese Merino sheep, Dorset sheep, Suffolk sheep, Texel, Hu sheep) in Xinjiang were analyzed by PCR-SSCP and PCR-RFLP method. The result showed:(1) 5 different sheep was midrange polymorphism at KAP1.3 loci, and PIC was between 0.25 and 0.5. Texel sheep not achieve genetic balance tested byχ2. (2) KAP81 loci Dorset sheep was high polymorphism (PIC>0.5), Texel sheep was low polymorphism (PIC<0.25), Chinese Merino sheep, Hu sheep and Suffolk sheep was midrange polymorphism (0.25
Secondly, study the relation of gene and wool fineness, strength and length breeding value
Study the relationship between Chinese Merino sheep wool traits breeding value and gene loci by multiple comparison and t tset, gene has relation of wool trait in different sheep, research in first part. The result showed that:(1)result of phenotypic value and individual breeding value: AC genotype of KAP81,AA genotype of KAP1.3,BC genotype of X611,could mark wool fineness; BC genotype of KAP81, AB genotype of KRT123 could mark wool strength; AA genotype of X611 could mark wool length. (2) The result of Study the genotype breeding value of 15 Sire sheep was: It is closer between genotype breeding value and phenotypic value of progeny .So it is convenience use homogamy to get high quality sheep.
引文
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