猪IGF-I基因及MyoG基因分子遗传多态性的研究
摘要
猪肌肉的生长速度直接关系到养猪生产的经济效益,肌肉生长主要指骨骼肌生长。肌肉生长潜力与肌纤维数量和肌纤维生长有关。肌细胞生成素(MyoG)基因是骨骼肌转录因子MyoD 家族的一员,在肌细胞生成过程中起着中心调控作用。胰岛素样生长因子I (IGF-I),是生长激素发挥促生长作用的重要调节因子。在肌细胞形成过程中,IGF-I 促进肌细胞分化。在骨骼肌生长过程中,IGF-I 调节骨骼肌的生长。IGF-I 通过诱导肌细胞生成素(MyoG)基因表达而刺激成肌细胞的终末端分化。胰岛素样生长因子I(IGF-I)和肌细胞生成素(MyoG)基因与猪肌肉的生长速度有关。
本研究对猪IGF-I 基因(外显子3 和外显子4)和MyoG 基因(5'-调控区及所有外显子区)做了单核苷酸多态性(SNPs)分析,并确立了最佳PCR-SSCP 分析条件。检测结果发现,在IGF-I 基因的外显子3 和外显子4,以及MyoG 基因外显子3、MyoG5'-1 和MyoG5'-3 共5 个座位上存在多态性。对存在多态性的5 个座位分别克隆和测序,测序结果显示在IGF-I 外显子3 有C→T,在IGF-I 外显子4 有C→T;在MyoG 基因外显子3 有T→C,在MyoG5'-1 有C→A(-473),C→A(-456),在MyoG5'-3 有A→T(-44)。6 个多态性位点文献中均未见报道。
猪MyoG 基因的5'-区存在多态性,对5'-区DNA 多态性位点的调控因子结合情况进行预测。结果显示,在-473、-44 位置碱基多态性引起了转录因子结合的变动,这些位置的碱基多态性可能会引起调控元件的变化。将存在多态性的外显子(IGF-I 基因外显子3和外显子4、MyoG 基因外显子3)不同纯合基因型个体的核苷酸序列分别翻译成氨基酸序列,然后进行氨基酸序列的同源性比较,结果显示,所有检测到的外显子序列上碱基的多态性并没有引起氨基酸序列的变化,说明这几个外显子上核苷酸序列的差异是遗传密码子的摇摆性所致。
对IGF-I基因和MyoG基因上所检测到的多态性片段分别做了基因频率和基因型频率统计。结果显示,在3 个外显子(IGF-I 基因外显子3 和外显子4 以及MyoG 基因外显子3)上,不同品种间基因频率的分布存在差异;而在MyoG5'-1 和MyoG5'-3 两个座位上,不同品种间基因频率的分布基本上是一致的。
对包括品种、场-年-季和IGF-I 基因以及MyoG 基因的基因型效应进行最小二乘分析,
The growth rate of porcine muscle is directly related to the economic benefits of pig production, and skeletal muscle growth is an important part of muscle growth in pig. Meat production capacity is related to muscle fiber numbers and growth. Myogenin belongs to the MyoD gene family and fulfills a key function in muscle differentiation by controlling the onset of myoblast fusion and the establishment of myofibers. Insulin-like growth factor I (IGF-I) is an important regulation factor for the functioning of growth hormon.
IGF-I stimulates differentiation through a myogenin pathway and functions in the regulation of muscle growth, it also stimulates terminal myogenic differentiation by induction of myogenin gene expression. Insulin –like growth factor I and myogenin are related to the growth rate of porcine skeletal muscle . IGF-I gene (exon3 and exon4) and MyoG gene (5’-regulation region and all exons) were amplified to identify the nucleotide diversity. IGF-I exon3, IGF-I exon4, MyoG exon3, MyoG5’-1 and MyoG5’-3 appeared polymorphic. The results of polymorphisms are IGF-I exon3 c→t, IGF-I exon4 c→t, MyoG exon3 t→c, MyoG5’-1 c→a(-473), c→a(-456), MyoG5’-3 a→t (-44).The five polymorphic loci were not reported in the document.
The potential elements and protein factors binding to the 5’-region of myogenin gene were predicted and different regulatory elements between polymorphisms of two loci (-473 and -44) were discovered. On these two sites, the changes of regulation elements were possibly made by the base changes. The alteration of a single base in exons (IGF-I exon3, IGF-I exon4 and MyoG exon3) are all silent mutation, no substitution of amino acid happened. The alteration of a single base in these exons were caused by the swinging of genetic coden.
Gene and genotype frequencies were calculated for the polymorphic loci in IGF-I gene and MyoG gene. Some differences of gene frequency between different breeds in IGF-I exon3, IGF-I exon4 and MyoG exon3 were discovered. No significant differences for the distribution of gene frequency were discovered between breeds on MyoG5’-1 and MyoG5’-3 loci.
The least square means of different IGF-I and MyoG genotype for growth traits were estimated with the fixed model. Backfat thickness and body weight at six months were influenced by the loci of IGF-I exon3 and 5’-1 in MyoG (P<0.05) respectively; Breed had significant effects on birth weight, weaning weight, body weight at six months at level of P<0.01 and on backfat thickness at level of P<0.05.
The results suggested that insulin-like growth factor I (IGF-I) gene and myogenin (MyoG) gene are potential genetic markers in the study on porcine muscle growth and could be used as markers in marker assisted selection to improve the efficiency and accuracy of selection.
本研究对猪IGF-I 基因(外显子3 和外显子4)和MyoG 基因(5'-调控区及所有外显子区)做了单核苷酸多态性(SNPs)分析,并确立了最佳PCR-SSCP 分析条件。检测结果发现,在IGF-I 基因的外显子3 和外显子4,以及MyoG 基因外显子3、MyoG5'-1 和MyoG5'-3 共5 个座位上存在多态性。对存在多态性的5 个座位分别克隆和测序,测序结果显示在IGF-I 外显子3 有C→T,在IGF-I 外显子4 有C→T;在MyoG 基因外显子3 有T→C,在MyoG5'-1 有C→A(-473),C→A(-456),在MyoG5'-3 有A→T(-44)。6 个多态性位点文献中均未见报道。
猪MyoG 基因的5'-区存在多态性,对5'-区DNA 多态性位点的调控因子结合情况进行预测。结果显示,在-473、-44 位置碱基多态性引起了转录因子结合的变动,这些位置的碱基多态性可能会引起调控元件的变化。将存在多态性的外显子(IGF-I 基因外显子3和外显子4、MyoG 基因外显子3)不同纯合基因型个体的核苷酸序列分别翻译成氨基酸序列,然后进行氨基酸序列的同源性比较,结果显示,所有检测到的外显子序列上碱基的多态性并没有引起氨基酸序列的变化,说明这几个外显子上核苷酸序列的差异是遗传密码子的摇摆性所致。
对IGF-I基因和MyoG基因上所检测到的多态性片段分别做了基因频率和基因型频率统计。结果显示,在3 个外显子(IGF-I 基因外显子3 和外显子4 以及MyoG 基因外显子3)上,不同品种间基因频率的分布存在差异;而在MyoG5'-1 和MyoG5'-3 两个座位上,不同品种间基因频率的分布基本上是一致的。
对包括品种、场-年-季和IGF-I 基因以及MyoG 基因的基因型效应进行最小二乘分析,
The growth rate of porcine muscle is directly related to the economic benefits of pig production, and skeletal muscle growth is an important part of muscle growth in pig. Meat production capacity is related to muscle fiber numbers and growth. Myogenin belongs to the MyoD gene family and fulfills a key function in muscle differentiation by controlling the onset of myoblast fusion and the establishment of myofibers. Insulin-like growth factor I (IGF-I) is an important regulation factor for the functioning of growth hormon.
IGF-I stimulates differentiation through a myogenin pathway and functions in the regulation of muscle growth, it also stimulates terminal myogenic differentiation by induction of myogenin gene expression. Insulin –like growth factor I and myogenin are related to the growth rate of porcine skeletal muscle . IGF-I gene (exon3 and exon4) and MyoG gene (5’-regulation region and all exons) were amplified to identify the nucleotide diversity. IGF-I exon3, IGF-I exon4, MyoG exon3, MyoG5’-1 and MyoG5’-3 appeared polymorphic. The results of polymorphisms are IGF-I exon3 c→t, IGF-I exon4 c→t, MyoG exon3 t→c, MyoG5’-1 c→a(-473), c→a(-456), MyoG5’-3 a→t (-44).The five polymorphic loci were not reported in the document.
The potential elements and protein factors binding to the 5’-region of myogenin gene were predicted and different regulatory elements between polymorphisms of two loci (-473 and -44) were discovered. On these two sites, the changes of regulation elements were possibly made by the base changes. The alteration of a single base in exons (IGF-I exon3, IGF-I exon4 and MyoG exon3) are all silent mutation, no substitution of amino acid happened. The alteration of a single base in these exons were caused by the swinging of genetic coden.
Gene and genotype frequencies were calculated for the polymorphic loci in IGF-I gene and MyoG gene. Some differences of gene frequency between different breeds in IGF-I exon3, IGF-I exon4 and MyoG exon3 were discovered. No significant differences for the distribution of gene frequency were discovered between breeds on MyoG5’-1 and MyoG5’-3 loci.
The least square means of different IGF-I and MyoG genotype for growth traits were estimated with the fixed model. Backfat thickness and body weight at six months were influenced by the loci of IGF-I exon3 and 5’-1 in MyoG (P<0.05) respectively; Breed had significant effects on birth weight, weaning weight, body weight at six months at level of P<0.01 and on backfat thickness at level of P<0.05.
The results suggested that insulin-like growth factor I (IGF-I) gene and myogenin (MyoG) gene are potential genetic markers in the study on porcine muscle growth and could be used as markers in marker assisted selection to improve the efficiency and accuracy of selection.
引文
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