不同猪种IGF1R基因扩增及SNPs的筛选
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摘要
以小型猪(巴马香猪46头)和大型猪(长白猪、大白猪、杜洛克猪,共198头)为试验对象,采用PCR产物直接测序法对胰岛素样生长因子1受体(IGF1R)外显子进行SNPs筛查,根据测序结果确定每个SNP位点的基因型,并通过Haploview软件对筛选到的SNPs进行连锁不平衡及单倍型分析。结果表明:在不同体型猪IGF1R基因外显子1~21上共筛选到9处同义SNPs位点,为G263C(rs338724264)、T17G(rs325909655)、C145T (rs337838116)、 T40C (rs326728191)、 C104T (rs319719983)、 C170T (rs338444971)、 C60T(rs327750836)、C126T(rs327750836)和C82T(rs339449791)。这9个SNPs在巴马香猪和大型猪中均为纯合子突变,且呈现完全连锁不平衡。此研究明确了不同体型猪IGF1R基因外显子区SNPs的基因型及由这些SNPs形成的优势单倍型,可为探究不同猪种生长发育过程中体尺存在差异的原因提供新数据。
A total of 46 miniature pigs( Bama miniature pigs) and 198 large pigs( Landrace,Yorkshire and Duroc pigs) were used as experimental subjects. SNPs in the exon region of the insulinlike growth factor 1 receptor( IGF1 R) were determined by direct sequencing of PCR products. The genotype of each SNP locus was determined based on the sequencing results,and the linkage disequilibrium and haplotype analysis of the selected SNPs were performed by Haploview software. The experimental results show that there are 9 synonymous SNPs in exon 1-21 of IGF1 R gene,which are as follows: G263 C( rs338724264), T17 G( rs325909655), C145 T( rs337838116), T40 C( rs326728191),C104 T( rs319719983),C170 T( rs338444971),C60 T( rs327750836),C126 T( rs327750836) and C82 T( rs339449791). The 9 SNPs showed complete linkage disequilibrium andformed a haplotype( P<0. 0001) in both Bama miniature pigs and large pigs. This study clarified the genotypes of SNPs in the exon regions of IGF1 R genes of different body type pigs and the dominant haplotypes formed by these SNPs providing new data for clarifying the reasons for the difference in body size during the growth and development of different pig breeds.
A total of 46 miniature pigs( Bama miniature pigs) and 198 large pigs( Landrace,Yorkshire and Duroc pigs) were used as experimental subjects. SNPs in the exon region of the insulinlike growth factor 1 receptor( IGF1 R) were determined by direct sequencing of PCR products. The genotype of each SNP locus was determined based on the sequencing results,and the linkage disequilibrium and haplotype analysis of the selected SNPs were performed by Haploview software. The experimental results show that there are 9 synonymous SNPs in exon 1-21 of IGF1 R gene,which are as follows: G263 C( rs338724264), T17 G( rs325909655), C145 T( rs337838116), T40 C( rs326728191),C104 T( rs319719983),C170 T( rs338444971),C60 T( rs327750836),C126 T( rs327750836) and C82 T( rs339449791). The 9 SNPs showed complete linkage disequilibrium andformed a haplotype( P<0. 0001) in both Bama miniature pigs and large pigs. This study clarified the genotypes of SNPs in the exon regions of IGF1 R genes of different body type pigs and the dominant haplotypes formed by these SNPs providing new data for clarifying the reasons for the difference in body size during the growth and development of different pig breeds.
引文
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[15]郝哲,孙永峰,隋玉健,等.鹅肌肉IGF-Ⅰ基因mRNA的表达[J].吉林农业大学学报,2012,34(2):184-188.
[16]史旭升,孙永峰,杨童奥,等.胰岛素样生长因子Ⅰ基因在鹅不同发育时期肌肉组织中的表达[J].吉林农业大学学报,2011,33(2):214-217.
[17]常新侠,王树迎,于学浩,等.济宁青山羊生后乳腺GHR和IGF-ⅠR的分布及其mRNA表达的发育性变化[J].中国兽医学报,2016,36(7):1234-1240.
[18] Lei M,Peng X,Zhou M,et al. Polymorphisms of the IGFIR gene and their genetic effects on chicken early growth and carcass traits[J]. Bmc Genetics,2008,9(1):70.
[19] Dao A,Sanou J,Mitchell S E,et al. Genetic diversity among INERA maize inbred lines with single nucleotide polymorphism(SNP)markers and their relationship with CIMMYT,IITA,and temperate lines[J]. Bmc Genetics,2014,15(1):127.
[20] Zhang K,Qin Z S,Liu J S,et al. Haplotype block partitioning and tag SNP selection using genotype data and their applications to association studies[J]. Genome Research,2004,14(5):908-916.
[2]商海涛,魏泓.我国小型猪品系资源状况初浅分析[J].中国实验动物学报,2007,15(1):70-75.
[3] Simianer H,K9hn F. Genetic management of the Gottingen Minipig population.[J]. Journal of Pharmacological&Toxicological Methods,2010,62(3):221.
[4] Deng J Z,Hao L L,Li M T,et al. Growth hormone and receptor gene mutations in Chinese Banna miniature pig[J]. Animal Cells&Systems,2011,15(4):310-314.
[5]郑鑫,刘国文,梁海涛,等.铜对猪原代肝细胞中类胰岛素生长因子及其结合蛋白的影响[J].吉林农业大学学报,2006,28(3):330-333.
[6] Mohn A,Marcovecchio M L,Giorgis T D,et al. An insulinlike growth factor-I receptor defect associated with short stature and impaired carbohydrate homeostasis in an italian pedigree[J]. Hormone Research in Paediatrics,2011,76(2):136-143.
[7] Klammt J,Pfffle R,Werner H,et al. IGF signaling defects as causes of growth failure and IUGR[J]. Trends in Endocrinology&Metabolism Tem,2008,19(6):197-205.
[8] Leroith D,Werner H,Beitnerjohnson D,et al. Molecular and cellular aspects of the insulin-like growth factor I receptor[J].Endocrine Reviews,2008,16(2):143-163.
[9] Ward C W,Menting J G,Lawrence M C. The insulin receptor changes conformation in unforeseen ways on ligand binding:sharpening the picture of insulin receptor activation[J]. Bioessays News&Reviews in Molecular Cellular&Developmental Biology,2013,35(11):945-954.
[10] Siddle K. Signalling by insulin and IGF receptors:supporting acts and new players[J]. Journal of Molecular Endocrinology,2011,47(1):1-10.
[11] Pierzchala M,Pareek C S,Urbanski P,et al. Study of the differential transcription in liver of growth hormone receptor(GHR),insulin-like growth factors(IGF1,IGF2)and insulin-like growth factor receptor(IGF1R)genes at different postnatal developmental ages in pig breeds[J]. Mol Biol Rep,2012,39(3):3055-3066.
[12] Dupont J,Holzenberger M. Biology of insulin-like growth factors in development[J]. Birth Defects Res C Embryo Today,2003,69(4):257-271.
[13]李常红,郝林琳,于浩,等.猪IGF-1及其受体基因部分组织表达规律的研究[J].中国兽医学报,2013,33(10):1568-1572.
[14]张昕,贾泓瑶,程云云,等.猪IGF-1R基因启动子区单核苷酸多态性检测分析[J].中国兽医学报,2014,34(10):1689-1692,1698.
[15]郝哲,孙永峰,隋玉健,等.鹅肌肉IGF-Ⅰ基因mRNA的表达[J].吉林农业大学学报,2012,34(2):184-188.
[16]史旭升,孙永峰,杨童奥,等.胰岛素样生长因子Ⅰ基因在鹅不同发育时期肌肉组织中的表达[J].吉林农业大学学报,2011,33(2):214-217.
[17]常新侠,王树迎,于学浩,等.济宁青山羊生后乳腺GHR和IGF-ⅠR的分布及其mRNA表达的发育性变化[J].中国兽医学报,2016,36(7):1234-1240.
[18] Lei M,Peng X,Zhou M,et al. Polymorphisms of the IGFIR gene and their genetic effects on chicken early growth and carcass traits[J]. Bmc Genetics,2008,9(1):70.
[19] Dao A,Sanou J,Mitchell S E,et al. Genetic diversity among INERA maize inbred lines with single nucleotide polymorphism(SNP)markers and their relationship with CIMMYT,IITA,and temperate lines[J]. Bmc Genetics,2014,15(1):127.
[20] Zhang K,Qin Z S,Liu J S,et al. Haplotype block partitioning and tag SNP selection using genotype data and their applications to association studies[J]. Genome Research,2004,14(5):908-916.