LTBP-1基因多态性与母猪繁殖性状关联分析
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摘要
目的 TGF-β信号通路在哺乳动物早期胚胎发育和初级卵泡、次级卵泡的转化过程中发挥着重要作用,而潜在性转化生长因子结合蛋白1(LTBP-1)是TGF-β的上调因子,直接决定了TGF-β的生物活性,进而间接影响哺乳动物的胚胎发育表现出不同的繁殖性状,但LTBP-1基因多态性对猪繁殖性能的影响还未见报道。方法本实验通过采用PCR-SSCP和CRS-PCR-RFLP技术,检测LTBP-1基因的两个SNP位点,对446头大白母猪进行分型并与繁殖性状关联分析。结果 LTBP-1基因C113404036G突变造成谷氨酰胺转变为组氨酸,检测到CC、CG、GG 3种基因型,各基因型大白母猪总仔数、活仔数、健仔数、死胎数、木乃伊胎数差异不显著,但CC型初生窝重显著高于CG型(P<0.05);大白猪LTBP-1基因A113356472G突变造成半胱氨酸转变为酪氨酸,检测到AA、AG 2种基因型,AG型母猪总产仔数显著高于AA型(P<0.05)、木乃伊胎数显著低于AA型(P<0.05)。结论 LTBP-1基因C113404036G和A113356475G两个错义突变位点,有望作为猪繁殖性状的分子标记。
Objective The TGF-β signaling pathway plays an important role in mammalian early embryonic development and transformation of primary follicles and secondary follicles,while latent transforming growth factor binding protein 1(LTBP-1) is an upregulating factor of TGF-β.The biological activity of TGF-β was determined,which indirectly affected the embryonic development of mammals and showed different reproductive traits.However,the effect of LTBP-1 gene polymorphism on reproductive performance of pigs has not been reported. Methods In this experiment,PCR-SSCP and CRS-PCR-RFLP techniques were used to detect the two SNP loci of the LTBP-1 gene found in the previous stage,and four hundred and forty-six large white sows of Zhengda Group pig farm were classified and associated with reproductive traits do polymorphism analysis. Results The results showed that the LTBP-1 gene C113404036 G mutation caused glutamine to be converted into histidine,and three genotypes of CC,CG and GG were detected.The total number of litters,live litters,litter size and number of stillbirths of each genotype of white sows There was no significant difference in the number of fetuses,but the CC type was significantly higher than the CG type(P<0.05).ConclusionThe LTBP-1 gene A113356472 G mutation in the Large White pig caused the conversion of cysteine to tyrosine,AA and AG two types were detected.The genotype,the total litter size of AG type sows was significantly higher than that of AA type(P<0.05),and the number of femmies was significantly lower than AA type(P<0.05).Conclusion We fand that the two missense mutation sites of LTBP-1 gene C113404036 G and A113356472 G con be used as molecular markers for repontuctive traits in pig.
Objective The TGF-β signaling pathway plays an important role in mammalian early embryonic development and transformation of primary follicles and secondary follicles,while latent transforming growth factor binding protein 1(LTBP-1) is an upregulating factor of TGF-β.The biological activity of TGF-β was determined,which indirectly affected the embryonic development of mammals and showed different reproductive traits.However,the effect of LTBP-1 gene polymorphism on reproductive performance of pigs has not been reported. Methods In this experiment,PCR-SSCP and CRS-PCR-RFLP techniques were used to detect the two SNP loci of the LTBP-1 gene found in the previous stage,and four hundred and forty-six large white sows of Zhengda Group pig farm were classified and associated with reproductive traits do polymorphism analysis. Results The results showed that the LTBP-1 gene C113404036 G mutation caused glutamine to be converted into histidine,and three genotypes of CC,CG and GG were detected.The total number of litters,live litters,litter size and number of stillbirths of each genotype of white sows There was no significant difference in the number of fetuses,but the CC type was significantly higher than the CG type(P<0.05).ConclusionThe LTBP-1 gene A113356472 G mutation in the Large White pig caused the conversion of cysteine to tyrosine,AA and AG two types were detected.The genotype,the total litter size of AG type sows was significantly higher than that of AA type(P<0.05),and the number of femmies was significantly lower than AA type(P<0.05).Conclusion We fand that the two missense mutation sites of LTBP-1 gene C113404036 G and A113356472 G con be used as molecular markers for repontuctive traits in pig.
引文
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[3]徐梦思.TGFβ-SMAD信号通路对猪颗粒细胞和繁殖性状的作用研究[D].石河子:石河子大学,2015.
[4]Miyazono K,Olofsson A,Colosetti P,et al.A role of the latent TGFβ1-binding protein in the assembly and secretion of TGF-β[J].EMBO J,1991,10(5):1091-1101.
[5]Saharinen J,Hyytiinen M,Taipale J,et al.Latent transforming growth factor-beta binding proteins(LTBPs)structural extracellular matrix proteins for targeting TGF-beta action[J].Cytokine Growth Factor Rev,1999,10(2):99-117.
[6]Weikkolainen K,Keskioja J,Koli K.Expression of latent TGF-beta binding protein LTBP-1 is hormaonally regulated in normak and transformed human lung fibroblasts[J].Growth Factors,2003,21:51-60.
[7]王威,赵扬玉,乔杰.LTBP-1在早孕胚胎停育患者绒毛蜕膜中的表达[J].山东医药,2009,49(44):23-25.Wei Wang,Yangyu Zhao,Jie Qiao.Expression of LTBP-1in the decidua of early pregnancy embryos[J].Shandong Medicine,2009,49(144):23-25.
[8]赵春江,李宁,邓学梅.应用创造酶切位点法检测单碱基突变[J].遗传,2003(3):327-329.Chunjiang Zhao,Ning Li,Xuemei Deng.Detecting single base mutations by creating a cleavage site[J].Genetics,2003(3):327-329.
[9]Jia Y,Gao G,Song H,et al.Low-protein diet fed to crossbredsows during pregnancy and lactation enhances myostatingene expression through epigenetic regulation in skeletalmuscle of weaning piglets[J].European Journal of Nutrition,2016,53(3):1307-1314
[10]Pangas S A.Regulation of the ovarian reserve by members of the transforming growth factor beta family[J].Molecular Reproduction and Development,2012,79(10):666-679.
[11]Pangas S A,Matzuk M M.Genetic models for transforming growth factorβsuperfamily signaling in ovarian follicle development[J].Mol Cell Endocrinol,2004(1-2),225:83-91.
[12]Derynck R,Aknurst R J,Balmain A.TGF-βsignaling in tumor suppression and cancer progression[J].Nature Genetics,2001,29(2):117-129.
[13]武艳萍.猪转化生生长因子β1基因多态性与猪产仔数关系的研究[D].北京:中国农业大学,2005.
[14]李海晶,傅金恋,王爱国,等.不同猪种TGFβRⅠ基因单核苷酸多态性分析[J].中国畜牧杂志,2008,44:13-15.Li H J,Fu J L,Wang A G,et al.Single nucleotide polymorphism analysis of TGF-βRI gene in different pig breeds[J].Chinese Journ of Animal Science 2008,44:13-15.
[15]李海晶.猪转化生长因子β受体Ⅰ基因多态性与猪产仔数关系的研究[D].北京:中国农业大学.2006.
[16]王嘉博.猪GSTM2、TGFβ1基因的多态性与生产性状相关性分析[D].哈尔滨:东北农业大学硕士论文,2010.
[17]Miyazono K,Olofsson A,Colosetti P,et al.A role of the latent TGFβ1-binding protein in the assembly and secretion of TGF-β[J].EMBO J,1991,10(5):1091-1101.
[18]Tatti O,Vehviinen P,Lehti K,et al.MT1-MMP releases latent TGFβ1 from endothelial cell extracellularmatrix via proteolytic processing of LTBP-1[J].Exp Cell Res,2008,314(13):2501-2514.
[2]郭晶,李新宇,李隐侠,等.湖羊TGF-β1基因特征、表达及其与排卵数的相关性分析[J].中国农业科学,2013,46(21):4586-4593Guo J,Li X Y,Li Y X,et al.Analysis of the relationship between TGF-β1 gene characteristics,expression and ovulation number in Hu sheep[J].Chinese Agricultural Sciences,2013,46(21):4586-4593
[3]徐梦思.TGFβ-SMAD信号通路对猪颗粒细胞和繁殖性状的作用研究[D].石河子:石河子大学,2015.
[4]Miyazono K,Olofsson A,Colosetti P,et al.A role of the latent TGFβ1-binding protein in the assembly and secretion of TGF-β[J].EMBO J,1991,10(5):1091-1101.
[5]Saharinen J,Hyytiinen M,Taipale J,et al.Latent transforming growth factor-beta binding proteins(LTBPs)structural extracellular matrix proteins for targeting TGF-beta action[J].Cytokine Growth Factor Rev,1999,10(2):99-117.
[6]Weikkolainen K,Keskioja J,Koli K.Expression of latent TGF-beta binding protein LTBP-1 is hormaonally regulated in normak and transformed human lung fibroblasts[J].Growth Factors,2003,21:51-60.
[7]王威,赵扬玉,乔杰.LTBP-1在早孕胚胎停育患者绒毛蜕膜中的表达[J].山东医药,2009,49(44):23-25.Wei Wang,Yangyu Zhao,Jie Qiao.Expression of LTBP-1in the decidua of early pregnancy embryos[J].Shandong Medicine,2009,49(144):23-25.
[8]赵春江,李宁,邓学梅.应用创造酶切位点法检测单碱基突变[J].遗传,2003(3):327-329.Chunjiang Zhao,Ning Li,Xuemei Deng.Detecting single base mutations by creating a cleavage site[J].Genetics,2003(3):327-329.
[9]Jia Y,Gao G,Song H,et al.Low-protein diet fed to crossbredsows during pregnancy and lactation enhances myostatingene expression through epigenetic regulation in skeletalmuscle of weaning piglets[J].European Journal of Nutrition,2016,53(3):1307-1314
[10]Pangas S A.Regulation of the ovarian reserve by members of the transforming growth factor beta family[J].Molecular Reproduction and Development,2012,79(10):666-679.
[11]Pangas S A,Matzuk M M.Genetic models for transforming growth factorβsuperfamily signaling in ovarian follicle development[J].Mol Cell Endocrinol,2004(1-2),225:83-91.
[12]Derynck R,Aknurst R J,Balmain A.TGF-βsignaling in tumor suppression and cancer progression[J].Nature Genetics,2001,29(2):117-129.
[13]武艳萍.猪转化生生长因子β1基因多态性与猪产仔数关系的研究[D].北京:中国农业大学,2005.
[14]李海晶,傅金恋,王爱国,等.不同猪种TGFβRⅠ基因单核苷酸多态性分析[J].中国畜牧杂志,2008,44:13-15.Li H J,Fu J L,Wang A G,et al.Single nucleotide polymorphism analysis of TGF-βRI gene in different pig breeds[J].Chinese Journ of Animal Science 2008,44:13-15.
[15]李海晶.猪转化生长因子β受体Ⅰ基因多态性与猪产仔数关系的研究[D].北京:中国农业大学.2006.
[16]王嘉博.猪GSTM2、TGFβ1基因的多态性与生产性状相关性分析[D].哈尔滨:东北农业大学硕士论文,2010.
[17]Miyazono K,Olofsson A,Colosetti P,et al.A role of the latent TGFβ1-binding protein in the assembly and secretion of TGF-β[J].EMBO J,1991,10(5):1091-1101.
[18]Tatti O,Vehviinen P,Lehti K,et al.MT1-MMP releases latent TGFβ1 from endothelial cell extracellularmatrix via proteolytic processing of LTBP-1[J].Exp Cell Res,2008,314(13):2501-2514.