绵羊STAT5a基因intron10和exon12遗传特征分析
|
摘要
【目的】研究绵羊STAT5a基因第10内含子和第12外显子的遗传多态性和群体遗传特征。【方法以2个多羔绵羊品种湖羊(140只)和多浪羊(144只)及单羔羊藏绵羊(217只)为研究对象,应用PCR-SSCP方法分析其STAT5a基因第10内含子和第12外显子的遗传多态性和群体遗传特征。【结果】绵羊STAT5a基因第10内含子发现13 210A>G突变,检测到3种基因型(AA、AG和GG)和2个等位基因(A和G);第12外显子上发现14 827A>G突变,该突变属于同义突变,检测到3种基因型(MM、MN和NN)和2个等位基因(M和N)。在绵羊STAT5a基因第10内含子和第12外显子中,杂合子AG和MN分别为优势基因型,基因型频率在多羔绵羊和单羔绵羊品种间差异极显著(P<0.01)。3个绵羊品种的多态信息含量均处于中度多态,藏绵羊的卡方值处于Hardy-Weinberg平衡状态,而湖羊和多浪羊偏离了Hardy-Weinberg平衡状态。【结论】绵羊STAT5a基因具有多态性,且基因型频率在不同产羔数品种间差异极显著(P<0.01)。
【Objective】This study investigated the genetic polymorphism and population genetic characteristics of intron 10 and exon 12 of STAT5 ain sheep.【Method】A total of 140 Hu sheep and 144 Duolang sheep of two polygamous sheep breeds as well as 217 Tibetan sheep of single lambs were selected and the PCR-SSCP was used to analyze the genetic polymorphism and population genetic characteristics of intron 10 and 12 exon of the STAT5 agene.【Results】The 13 210 A>G mutation was found in the intron 10 of STAT5 agene including three genotypes(AA,AG and GG)and two alleles(A and G).The 14 827 A>G synonymous mutation including three genotypes(MM,MN and NN)and two alleles(M and N)were detected in the exon 12.Heterozygous AG and MN were dominant genotypes in intron 10 and exon 12.The frequencies of genotypes were significantly different among multiple lambs and single lambs(P<0.01).The polymorphic information of the three sheep breeds was all moderately polymorphic.The Chi-square value of Tibetan sheep was in Hardy-Weinberg equilibrium,while that of Hu sheep and Duolang sheep deviated from Hardy-Weinberg equilibrium.【Conclusion】STAT5 awas polymorphic and the genotype frequencies of lambs were significantly different among different litter sizes(P<0.01).
【Objective】This study investigated the genetic polymorphism and population genetic characteristics of intron 10 and exon 12 of STAT5 ain sheep.【Method】A total of 140 Hu sheep and 144 Duolang sheep of two polygamous sheep breeds as well as 217 Tibetan sheep of single lambs were selected and the PCR-SSCP was used to analyze the genetic polymorphism and population genetic characteristics of intron 10 and 12 exon of the STAT5 agene.【Results】The 13 210 A>G mutation was found in the intron 10 of STAT5 agene including three genotypes(AA,AG and GG)and two alleles(A and G).The 14 827 A>G synonymous mutation including three genotypes(MM,MN and NN)and two alleles(M and N)were detected in the exon 12.Heterozygous AG and MN were dominant genotypes in intron 10 and exon 12.The frequencies of genotypes were significantly different among multiple lambs and single lambs(P<0.01).The polymorphic information of the three sheep breeds was all moderately polymorphic.The Chi-square value of Tibetan sheep was in Hardy-Weinberg equilibrium,while that of Hu sheep and Duolang sheep deviated from Hardy-Weinberg equilibrium.【Conclusion】STAT5 awas polymorphic and the genotype frequencies of lambs were significantly different among different litter sizes(P<0.01).
引文
[1]James E,Darnell J,Lan M.Jak-STAT pathways and trnascriptional activation in response to IFNs and other extracellular signaling proteins[J].Science,1994,264(5164):1415-1421.
[2]Schmittney M,Doppler W,Ball R K,et al.Beta-casein gene promoter activity is regulated by the hormone-mediated relief of transcriptional repression and a mammary-gland-specific nuclear factor[J].Molecular and Cellular Biology,1991,11(7):3745-3755.
[3]Miyoshi K,Shillingford J M,Smith G H,et al.Signal transducer and activator of transcription(Stat)5controls the proliferation and differentiation of mammary alveolar epithelium[J].Journal of Cell Bilolgy,2001,155(4):531-542.
[4]Bole-Feysot C,Goffin V,Edery M,et al.Prolactin(PRL)and its receptor:actions,signal transduction pathways and phenotypes observed in PRL receptor knockout mice[J].Endocrine Reviews,1998,19(3):225-268.
[5]Boutinaud M,Jammes H.Growth hormone increases Stat5and Stat1expression in lactating goat mammary gland:a specific effect compared to milking frequency[J].Domestic Animal Endocrinology,2004,27(4):363-378.
[6]Kaplan M H.STAT signaling in inflammation[J].JAK-STAT,2013,2(1):e24198.
[7]Richard A J,Stephens J M.Emerging roles of JAK-STAT signaling pathways in adipocytes[J].Trends in Endocrinology&Metabolism Tem,2011,22(8):325-332.
[8]吴贤锋.山羊STAT3和STAT5A基因SNPs、mRNA表达及甲基化对生产性能的影响[D].陕西杨凌:西北农林科技大学,2015.Wu X F.SNPs,mRNA expression and DNA methylation of STAT3and STAT5Agenes and their influences on production traits in goats[D].Yangling,Shaanxi:Northwest A&F University,2015.
[9]Brym P,Kaminski S.New SSCP polymorphism with in bovine STAT5A gene and its associations with milk performance traits[J].Journal of Applied Genetics,2004,45(4):445-452.
[10]何峰,孙东晓,余英,等.荷斯坦奶牛STAT5A基因的SNPs检测及其与产奶性状关联分析[J].畜牧兽医学报,2007,38(4):326-331.He F,Sun D X,Yu Y,et al.SNPs detection of STAT5Agene and association with milk production traits in holstein cattle[J].Acta Veterinaria et Zootechnica Sinica,2007,38(4):326-331.
[11]鲍斌,房兴堂,陈宏,等.中国荷斯坦牛STAT5A基因多态性与泌乳性状的相关分析[J].中国农业科学,2008,41(6):1872-1878.Bao B,Fang X T,Chen H,et al.Polymorphisms of STAT5Agene and its association with milk performance traits in chinese holstein cattle[J].Scientia Agricultura Sinica,2008,41(6):1872-1878.
[12]季敏,刘学洪,余长林,等.槟榔江水牛STAT5A基因多态性及其与产奶性状的关联性研究[J].中国牛业科学,2013,39(4):29-35.Ji M,Liu X H,Yu C L,et al.Association of STAT5A polymorphism and milk production traits in binlangjiang buffalo[J].China Cattle Science,2013,39(4):29-35.
[13]刘园峰,王桂芝,李秋梅,等.崂山奶山羊LEP和STAT5a基因对泌乳和生长性状的影响[J].中国农业科学,2013,46(18):3946-3954.Liu Y F,Wang G Z,Li Q M,et al.The effect of LEP and STAT5agenes on milk and growth traits of laoshan dairy goat[J].Scientia Agricultura Sinica,2013,46(18):3946-3954.
[14]谢海强,孙岩岩,龚俞,等.贵州黑山羊STAT5A基因多态性及其与生长性状关联分析[J].生物技术,2014(4):79-83.Xie H Q,Sun Y Y,Gong Y,et al.Association analysis between polymorphism of STAT5A gene and growth traits in guizhou black goat[J].Biotechnology,2014(4):79-83.
[15]Li H,Zheng H L.Signal transducer and activator of transcription 5ainhibited by pimozide may regulate survival of goat mammary gland epithelial cells by regulating parathyroid hormone-related protein[J].Gene,2014,51(2):279-289.
[16]郝志云,王继卿,胡江,等.绵羊STAT5a基因的组织表达及核苷酸序列变异分析[J].农业生物技术学报,2018,26(1):96-103.Hao Z Y,Wang J Q,Hu J,et al.The tissue expression of the sheep(ovis aries)STAT5agene and the analysis of nucleotide sequence variation[J].Journal of Agricultural Biotechnology,2018,26(1):96-103.
[17]Zhou H T,Hickford J G,Fang Q.A two-step procedure for extracting genomic DNA from dried blood spots on filter paper for polymerase chain reaction amplification[J].Analytical Biochemistry,2006,354(1):159-161.
[18]Byun S,Fang Q,Zhou H T,et al.An effective method for silver-staining DNA in large number of polyacrylamide gels[J].Analytical Biochemistry,2009,385(1):174-175.
[19]Gong H,Zhou H T,Hickford J G.Diversity of the glycine/tyrosine-rich keratin-associated protein 6gene(KAP6)family in sheep[J].Molecular Biology Reports,2011,38(1):31-35.
[20]Park Y W,Juárez M,Ramos M,et al.Physico-chemical characteristics of goat and sheep milk[J].Small Ruminant Research,2007,68(1/2):88-113.
[21]Kaleri R.Heritability estimates for some performances traits of baluchi sheep[J].Journal of Basic&Applied Sciences,2017,13,114-116.
[22]Geenty K G.Effects of weaning age on export lamb production[J].Proceedings of the New Zealand Society of Animal Production,1979,39:202-210.
[23]赵有璋.中国养羊学[M].北京:中国农业出版社,2013.Zhao Y Z.Chinese sheep science[M].Beijing:China Agriculture Press,2013.
[24]李庆章.奶牛乳腺发育与泌乳生物学[M].北京:科学出版社,2014.Li Q Z.Mammary gland development and lactation biology of daily cow[M].Beijing:Science Press,2014.
[25]Nott A,Meislin S H,Moore M J.A quantitative analysis of intron effects on mammalian gene expression[J].RNA-a Publication of the RNA Society,2003,9(5):607-617.
[26]Wang J,Zhou H,Rachel H J,et al.Variation in the ovine MYF5gene and its effect on carcass lean meat yield in New Zealand Romney sheep[J].Meat Science,2017,131:146-151.
[27]Gam R,Shah P,Crossland R E,et al.Genetic association of hematopoietic stem cell transplantation outcome beyond histocompatibility genes[J].Front Immunol,2017,8(899/905):380.
[28]Cases E,White S N,Riler D G,et al.Assessment of single nucleotide polymorphisms in genes residing on chromosomes 14and 29for association with carcass composition traits in Bos indicus cattle[J].Journal of Animal Science,2005,83(1):13-19.
[29]王以婷,杨卫红,赵云龙,等.CYP3A4内含子10对CYP3A4基因表达的增强子作用[J].昆明医科大学学报,2017,38(1):13-17.Wang Y T,Yang W H,Zhao Y L,et al.The enhancer role of CYP3A4intron 10in CYP3A4gene expression[J].Journal of Kunming Medical University,2017,38(1):13-17.
[30]Chang K C.Critical regulatory domains in intron 2of porcine sarcomeric myosin heavy chain gene[J].Journal of Muscle Research and Cell Motility,2000,21:451-461.
[31]Chen S,Li K,Cao W,et al.Codon-resolution analysis reveals a direct and context-dependent impact of individual synonymous mutations on mRNA level[J].Molecular Biology&Evolution,2017,34(11):2944-2958.
[32]Seligmann H,Warthi G.Genetic code optimization for cotranslational protein folding:codon directional asymmetry correlates with antiparallel betasheets,trna synthetase classes[J].Computational&Structural Biotechnology Journal,2017,15:412-424.
[33]Escudero J A,Nivina A,Cambray G,et al.Recoding of synonymous genes to expand evolutionary landscapes requires control of secondary structure affecting translation[J].Biotechnology&Bioengineering,2018,115(1):184-191.
[2]Schmittney M,Doppler W,Ball R K,et al.Beta-casein gene promoter activity is regulated by the hormone-mediated relief of transcriptional repression and a mammary-gland-specific nuclear factor[J].Molecular and Cellular Biology,1991,11(7):3745-3755.
[3]Miyoshi K,Shillingford J M,Smith G H,et al.Signal transducer and activator of transcription(Stat)5controls the proliferation and differentiation of mammary alveolar epithelium[J].Journal of Cell Bilolgy,2001,155(4):531-542.
[4]Bole-Feysot C,Goffin V,Edery M,et al.Prolactin(PRL)and its receptor:actions,signal transduction pathways and phenotypes observed in PRL receptor knockout mice[J].Endocrine Reviews,1998,19(3):225-268.
[5]Boutinaud M,Jammes H.Growth hormone increases Stat5and Stat1expression in lactating goat mammary gland:a specific effect compared to milking frequency[J].Domestic Animal Endocrinology,2004,27(4):363-378.
[6]Kaplan M H.STAT signaling in inflammation[J].JAK-STAT,2013,2(1):e24198.
[7]Richard A J,Stephens J M.Emerging roles of JAK-STAT signaling pathways in adipocytes[J].Trends in Endocrinology&Metabolism Tem,2011,22(8):325-332.
[8]吴贤锋.山羊STAT3和STAT5A基因SNPs、mRNA表达及甲基化对生产性能的影响[D].陕西杨凌:西北农林科技大学,2015.Wu X F.SNPs,mRNA expression and DNA methylation of STAT3and STAT5Agenes and their influences on production traits in goats[D].Yangling,Shaanxi:Northwest A&F University,2015.
[9]Brym P,Kaminski S.New SSCP polymorphism with in bovine STAT5A gene and its associations with milk performance traits[J].Journal of Applied Genetics,2004,45(4):445-452.
[10]何峰,孙东晓,余英,等.荷斯坦奶牛STAT5A基因的SNPs检测及其与产奶性状关联分析[J].畜牧兽医学报,2007,38(4):326-331.He F,Sun D X,Yu Y,et al.SNPs detection of STAT5Agene and association with milk production traits in holstein cattle[J].Acta Veterinaria et Zootechnica Sinica,2007,38(4):326-331.
[11]鲍斌,房兴堂,陈宏,等.中国荷斯坦牛STAT5A基因多态性与泌乳性状的相关分析[J].中国农业科学,2008,41(6):1872-1878.Bao B,Fang X T,Chen H,et al.Polymorphisms of STAT5Agene and its association with milk performance traits in chinese holstein cattle[J].Scientia Agricultura Sinica,2008,41(6):1872-1878.
[12]季敏,刘学洪,余长林,等.槟榔江水牛STAT5A基因多态性及其与产奶性状的关联性研究[J].中国牛业科学,2013,39(4):29-35.Ji M,Liu X H,Yu C L,et al.Association of STAT5A polymorphism and milk production traits in binlangjiang buffalo[J].China Cattle Science,2013,39(4):29-35.
[13]刘园峰,王桂芝,李秋梅,等.崂山奶山羊LEP和STAT5a基因对泌乳和生长性状的影响[J].中国农业科学,2013,46(18):3946-3954.Liu Y F,Wang G Z,Li Q M,et al.The effect of LEP and STAT5agenes on milk and growth traits of laoshan dairy goat[J].Scientia Agricultura Sinica,2013,46(18):3946-3954.
[14]谢海强,孙岩岩,龚俞,等.贵州黑山羊STAT5A基因多态性及其与生长性状关联分析[J].生物技术,2014(4):79-83.Xie H Q,Sun Y Y,Gong Y,et al.Association analysis between polymorphism of STAT5A gene and growth traits in guizhou black goat[J].Biotechnology,2014(4):79-83.
[15]Li H,Zheng H L.Signal transducer and activator of transcription 5ainhibited by pimozide may regulate survival of goat mammary gland epithelial cells by regulating parathyroid hormone-related protein[J].Gene,2014,51(2):279-289.
[16]郝志云,王继卿,胡江,等.绵羊STAT5a基因的组织表达及核苷酸序列变异分析[J].农业生物技术学报,2018,26(1):96-103.Hao Z Y,Wang J Q,Hu J,et al.The tissue expression of the sheep(ovis aries)STAT5agene and the analysis of nucleotide sequence variation[J].Journal of Agricultural Biotechnology,2018,26(1):96-103.
[17]Zhou H T,Hickford J G,Fang Q.A two-step procedure for extracting genomic DNA from dried blood spots on filter paper for polymerase chain reaction amplification[J].Analytical Biochemistry,2006,354(1):159-161.
[18]Byun S,Fang Q,Zhou H T,et al.An effective method for silver-staining DNA in large number of polyacrylamide gels[J].Analytical Biochemistry,2009,385(1):174-175.
[19]Gong H,Zhou H T,Hickford J G.Diversity of the glycine/tyrosine-rich keratin-associated protein 6gene(KAP6)family in sheep[J].Molecular Biology Reports,2011,38(1):31-35.
[20]Park Y W,Juárez M,Ramos M,et al.Physico-chemical characteristics of goat and sheep milk[J].Small Ruminant Research,2007,68(1/2):88-113.
[21]Kaleri R.Heritability estimates for some performances traits of baluchi sheep[J].Journal of Basic&Applied Sciences,2017,13,114-116.
[22]Geenty K G.Effects of weaning age on export lamb production[J].Proceedings of the New Zealand Society of Animal Production,1979,39:202-210.
[23]赵有璋.中国养羊学[M].北京:中国农业出版社,2013.Zhao Y Z.Chinese sheep science[M].Beijing:China Agriculture Press,2013.
[24]李庆章.奶牛乳腺发育与泌乳生物学[M].北京:科学出版社,2014.Li Q Z.Mammary gland development and lactation biology of daily cow[M].Beijing:Science Press,2014.
[25]Nott A,Meislin S H,Moore M J.A quantitative analysis of intron effects on mammalian gene expression[J].RNA-a Publication of the RNA Society,2003,9(5):607-617.
[26]Wang J,Zhou H,Rachel H J,et al.Variation in the ovine MYF5gene and its effect on carcass lean meat yield in New Zealand Romney sheep[J].Meat Science,2017,131:146-151.
[27]Gam R,Shah P,Crossland R E,et al.Genetic association of hematopoietic stem cell transplantation outcome beyond histocompatibility genes[J].Front Immunol,2017,8(899/905):380.
[28]Cases E,White S N,Riler D G,et al.Assessment of single nucleotide polymorphisms in genes residing on chromosomes 14and 29for association with carcass composition traits in Bos indicus cattle[J].Journal of Animal Science,2005,83(1):13-19.
[29]王以婷,杨卫红,赵云龙,等.CYP3A4内含子10对CYP3A4基因表达的增强子作用[J].昆明医科大学学报,2017,38(1):13-17.Wang Y T,Yang W H,Zhao Y L,et al.The enhancer role of CYP3A4intron 10in CYP3A4gene expression[J].Journal of Kunming Medical University,2017,38(1):13-17.
[30]Chang K C.Critical regulatory domains in intron 2of porcine sarcomeric myosin heavy chain gene[J].Journal of Muscle Research and Cell Motility,2000,21:451-461.
[31]Chen S,Li K,Cao W,et al.Codon-resolution analysis reveals a direct and context-dependent impact of individual synonymous mutations on mRNA level[J].Molecular Biology&Evolution,2017,34(11):2944-2958.
[32]Seligmann H,Warthi G.Genetic code optimization for cotranslational protein folding:codon directional asymmetry correlates with antiparallel betasheets,trna synthetase classes[J].Computational&Structural Biotechnology Journal,2017,15:412-424.
[33]Escudero J A,Nivina A,Cambray G,et al.Recoding of synonymous genes to expand evolutionary landscapes requires control of secondary structure affecting translation[J].Biotechnology&Bioengineering,2018,115(1):184-191.