BMP-15基因外显子Ⅰ在三种绵羊中的多态性及与繁殖力相关性分析
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摘要
为研究绵羊繁殖力与绵羊BMP-15 (bone morphogenetic protein 15)基因的多态位点关系,并寻找调控绵羊繁殖力的分子标记,以甘肃高山细毛羊、蒙古羊、小尾寒羊三种绵羊为研究对象,采用PCR-SSCP技术与DNA碱基测序相结合的方法,检测该基因第一外显子67、92两个位点在上述三个不同品种绵羊中是否存在FecX~H (Q23→Ter)和FecX~I (V31→D)突变,同时根据检测结果与其繁殖力做相关性分析。结果表明,在三种绵羊中既未检测出与Inverdale绵羊相同的FecX~I突变,也未检测出与Romney绵羊相同的FecX~H突变,因此推测BMP-15基因中影响Romney与Inverdale绵羊高繁殖力的突变位点对以上三种绵羊均无显著影响。
To study the relationship between fecundity in sheep and sheep BMP-15 gene polymorphism sites, and to seek for molecular markers to regulate the fecundity of sheep, we took Gansu alpine fine-wool sheep, Mongolian sheep, small tail Han sheep as the objects of this study and used the way of combination of PCR-SSCP technology and DNA base sequencing to detected whether there were FecX~ H(Q23→Ter) and FecX~ I(V31→D) mutations at the two sites of the first exon 67, 92 of the gene in the three different breeds of sheep, then we analyzed the correlation between the test results and the fecundity.The results showed that neither the same FecX~ I mutation as the Inverdale sheep nor the same FecX~ Hmutation as the Romney sheep were detected in the three kinds of sheep. Therefore, it was speculated that the mutation sites affecting the high fecundity of Romney and Inverdale sheep in BMP-15 gene might have no significant effect on the above three kinds of sheep.
To study the relationship between fecundity in sheep and sheep BMP-15 gene polymorphism sites, and to seek for molecular markers to regulate the fecundity of sheep, we took Gansu alpine fine-wool sheep, Mongolian sheep, small tail Han sheep as the objects of this study and used the way of combination of PCR-SSCP technology and DNA base sequencing to detected whether there were FecX~ H(Q23→Ter) and FecX~ I(V31→D) mutations at the two sites of the first exon 67, 92 of the gene in the three different breeds of sheep, then we analyzed the correlation between the test results and the fecundity.The results showed that neither the same FecX~ I mutation as the Inverdale sheep nor the same FecX~ Hmutation as the Romney sheep were detected in the three kinds of sheep. Therefore, it was speculated that the mutation sites affecting the high fecundity of Romney and Inverdale sheep in BMP-15 gene might have no significant effect on the above three kinds of sheep.
引文
Davis G.H.,Bruce G.D.,and Dodds K.G.,2001,Ovulation rate and litter size of prolific Inverdale(FecXI)and Hanna(FecXH)sheep,Biology of Reproduction,14(2):1-11
Davis G.H.,Galloway S.M.,Ross I.K.,Gregan S.M.,Ward J.,and Nimbkar B.V.,2002,DNA tests in prolific sheep from eight countries provide new evidence on origin of the booroola(fecb)mutation,Biology of Reproduction,66(6):1869
Demars J.,Fabre S.,Sarry J.,Rossetti R.,Gilbert H.,Persani L.,Tosser-Klopp G.,Mulsant P.,Nowak Z.,Drobik W.,Martyniuk E.,and Bodin L.,2013,Genome-Wide Association Studies Identify Two Novel BMP15 Mutations Responsible for an Atypical Hyperprolificacy Phenotype in Sheep,PLo SGenetics,9(4):e1003482
Dube J.L,Wang P.,Elvin J.,Lyons K.M.,Celeste A.J.,and Matzuk M.M.,1998,The bone morphogenetic protein 15gene is X-linked and expressed in oocytes,Molecular Endocrinology,12(12):1809-1817
Galloway S.M.,Mcnatty K.P.,Cambridge L.M.,Laitinen M.P.,and Juengel J.L.,2000,Mutations in an oocyte-derived growth factor gene(bmp15)cause increased ovulation rate and infertility in a dosage-sensitive manner,Nature Genetics,25(3):279-280
Hanrahan J.P,Gregan S.M,Mulsant P.,Mullen M.,and Davis G.H.,2004,Mutations in the genes for oocyte-derived growth factors GDF9 and BMP15 are associated with both increased ovulation rate and sterility in Cambridge and Belclare sheep(Ovis aries),Biology of Reproduction,70(4):900
Juengel J.L.,Hudson N.L.,Heath D.A.,Smith P.,and Reader K.L.,2002,Growth differentiation factor 9 and bone morphogenetic protein 15 are essential for ovarian follicular development in sheep,Biology of Reproduction,67(6):1777-1778
Liu S.F.,Jiang Y.L.,and Du L.X.,2003,Studies of BMPR-IB and BMP15 as candidate genes for fecundity in little tailed han sheep,Yichuan Xuebao(Acta Genetica Sinica),30(8):755-760(柳淑芳,姜运良,杜立新,2003,BMPR-IB和BMP15基因作为小尾寒羊多胎性能候选基因的研究,遗传学报,30(8):755-760)
Otsuka F.,and Shimasaki S.,2002,A negative feedback system between oocyte bone morphogenetic protein 15 and granulosa cell kit ligand:its role in regulating granulosa cell mitosis,Proceedings of the National Academy of Sciences of the United States of America,99(12):8060-8065
Qin Y.F.,Li W.,Chen Y.S.,and He Z.Y.,2017,Advances in BMP15 gene research,Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),25(2):324-334(覃玉凤,李维,陈瑶生,何祖勇,2017,BMP15基因研究进展,农业生物技术学报,25(2):324-334)
Sun R.Z.,Cheng L.,Shan Z.Y.,Lliu C.J.,Zhang J.X.,Lei L.,and Liu Z.H.,2009.,Role of GDF-9 and BMP-15 during folliculogenesis,Xiandai Shengwu Yixue Jinzhan(Progress in Modern Biomedicine),9(17):3351-3353(孙瑞珍,程琳,单智焱,刘春佳,张家新,雷蕾,刘忠华,2009,卵泡发生过程中GDF-9和BMP-15(GDF-9B)的作用,现代生物医学进展,9(17):3351-3353)
Tian X.,Sun H.X.,and Wang Y.J.,2009,Genetic polymorphism of bmpr-ib gene and effect on litter size in three sheep breeds,Xibei Nonglin Keji Daxue Xuebao(Ziran Kexue Ban)(Journal of Northwest A&F University:Natural Science Edition),37(11):31-36(田秀娥,孙红霞,王永军,2009,3个绵羊群体bmpr-ib基因的遗传多态性及其对产羔数的影响,西北农林科技大学学报(自然科学版),37(11):31-36)
Wang M.,Shi X.,Huang X.,Liu X.F.,Qin Y.F.,and Liu X.H.,2017,Improving gene targeting efficiency on the porcine bmp15gene mediated by crispr/cas9 by using the rgs surrogate reporter system,Yichuan(Hereditas),39(1):48-55(王敏,石翾,黄翔,刘小凤,覃玉凤,刘小红,2017,应用rgs双荧光替代性报告载体提高crispr/cas9对猪bmp15基因的打靶效率,遗传,39(1):48-55)
Wang Q.G.,Zhong F.G.,Li H.,Wang X.H.,Liu S.R.,Chen X.J.,and Gan S.Q.,2005,Detection of major gene on litter size in sheep,Yichuan(Hereditas),27(1):80-84(王启贵,钟发刚,李辉,王新华,刘守仁,陈晓军,甘尚泉,2005,绵羊产羔性状主效基因检测研究,遗传,27(1):80-84)
Yan C.,Wang P.,DeMayo J.,DeMayo F.J.,Elvin J.A.,Carino C.,Prasad S.V.,Skinner S.S.,Dunbar B.S.,Dube J.L.,Celeste A.J.,and Matzuk M.M.,2001,Synergistic roles of bone morphogenetic protein 15 and growth differentiation factor 9 in ovarian function,Molecular Endocrinology,15(6):854-66
Yi S.E.,Lapolt P.S.,Yoon B.S.,Chen J.Y.,Lu J.K.,and Lyons K.M.,2001,The type I bmp receptor bmprib is essential for female reproductive function,Proceedings of the National A-cademy of Sciences of the United States of America,98(14):7994-7999
Davis G.H.,Galloway S.M.,Ross I.K.,Gregan S.M.,Ward J.,and Nimbkar B.V.,2002,DNA tests in prolific sheep from eight countries provide new evidence on origin of the booroola(fecb)mutation,Biology of Reproduction,66(6):1869
Demars J.,Fabre S.,Sarry J.,Rossetti R.,Gilbert H.,Persani L.,Tosser-Klopp G.,Mulsant P.,Nowak Z.,Drobik W.,Martyniuk E.,and Bodin L.,2013,Genome-Wide Association Studies Identify Two Novel BMP15 Mutations Responsible for an Atypical Hyperprolificacy Phenotype in Sheep,PLo SGenetics,9(4):e1003482
Dube J.L,Wang P.,Elvin J.,Lyons K.M.,Celeste A.J.,and Matzuk M.M.,1998,The bone morphogenetic protein 15gene is X-linked and expressed in oocytes,Molecular Endocrinology,12(12):1809-1817
Galloway S.M.,Mcnatty K.P.,Cambridge L.M.,Laitinen M.P.,and Juengel J.L.,2000,Mutations in an oocyte-derived growth factor gene(bmp15)cause increased ovulation rate and infertility in a dosage-sensitive manner,Nature Genetics,25(3):279-280
Hanrahan J.P,Gregan S.M,Mulsant P.,Mullen M.,and Davis G.H.,2004,Mutations in the genes for oocyte-derived growth factors GDF9 and BMP15 are associated with both increased ovulation rate and sterility in Cambridge and Belclare sheep(Ovis aries),Biology of Reproduction,70(4):900
Juengel J.L.,Hudson N.L.,Heath D.A.,Smith P.,and Reader K.L.,2002,Growth differentiation factor 9 and bone morphogenetic protein 15 are essential for ovarian follicular development in sheep,Biology of Reproduction,67(6):1777-1778
Liu S.F.,Jiang Y.L.,and Du L.X.,2003,Studies of BMPR-IB and BMP15 as candidate genes for fecundity in little tailed han sheep,Yichuan Xuebao(Acta Genetica Sinica),30(8):755-760(柳淑芳,姜运良,杜立新,2003,BMPR-IB和BMP15基因作为小尾寒羊多胎性能候选基因的研究,遗传学报,30(8):755-760)
Otsuka F.,and Shimasaki S.,2002,A negative feedback system between oocyte bone morphogenetic protein 15 and granulosa cell kit ligand:its role in regulating granulosa cell mitosis,Proceedings of the National Academy of Sciences of the United States of America,99(12):8060-8065
Qin Y.F.,Li W.,Chen Y.S.,and He Z.Y.,2017,Advances in BMP15 gene research,Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),25(2):324-334(覃玉凤,李维,陈瑶生,何祖勇,2017,BMP15基因研究进展,农业生物技术学报,25(2):324-334)
Sun R.Z.,Cheng L.,Shan Z.Y.,Lliu C.J.,Zhang J.X.,Lei L.,and Liu Z.H.,2009.,Role of GDF-9 and BMP-15 during folliculogenesis,Xiandai Shengwu Yixue Jinzhan(Progress in Modern Biomedicine),9(17):3351-3353(孙瑞珍,程琳,单智焱,刘春佳,张家新,雷蕾,刘忠华,2009,卵泡发生过程中GDF-9和BMP-15(GDF-9B)的作用,现代生物医学进展,9(17):3351-3353)
Tian X.,Sun H.X.,and Wang Y.J.,2009,Genetic polymorphism of bmpr-ib gene and effect on litter size in three sheep breeds,Xibei Nonglin Keji Daxue Xuebao(Ziran Kexue Ban)(Journal of Northwest A&F University:Natural Science Edition),37(11):31-36(田秀娥,孙红霞,王永军,2009,3个绵羊群体bmpr-ib基因的遗传多态性及其对产羔数的影响,西北农林科技大学学报(自然科学版),37(11):31-36)
Wang M.,Shi X.,Huang X.,Liu X.F.,Qin Y.F.,and Liu X.H.,2017,Improving gene targeting efficiency on the porcine bmp15gene mediated by crispr/cas9 by using the rgs surrogate reporter system,Yichuan(Hereditas),39(1):48-55(王敏,石翾,黄翔,刘小凤,覃玉凤,刘小红,2017,应用rgs双荧光替代性报告载体提高crispr/cas9对猪bmp15基因的打靶效率,遗传,39(1):48-55)
Wang Q.G.,Zhong F.G.,Li H.,Wang X.H.,Liu S.R.,Chen X.J.,and Gan S.Q.,2005,Detection of major gene on litter size in sheep,Yichuan(Hereditas),27(1):80-84(王启贵,钟发刚,李辉,王新华,刘守仁,陈晓军,甘尚泉,2005,绵羊产羔性状主效基因检测研究,遗传,27(1):80-84)
Yan C.,Wang P.,DeMayo J.,DeMayo F.J.,Elvin J.A.,Carino C.,Prasad S.V.,Skinner S.S.,Dunbar B.S.,Dube J.L.,Celeste A.J.,and Matzuk M.M.,2001,Synergistic roles of bone morphogenetic protein 15 and growth differentiation factor 9 in ovarian function,Molecular Endocrinology,15(6):854-66
Yi S.E.,Lapolt P.S.,Yoon B.S.,Chen J.Y.,Lu J.K.,and Lyons K.M.,2001,The type I bmp receptor bmprib is essential for female reproductive function,Proceedings of the National A-cademy of Sciences of the United States of America,98(14):7994-7999