[1]金雯雯,王德迪,潘增祥,等.湖羊和巴什拜羊BMP15基因c.-1760C>A变异与启动子区活性的关系[J].南京农业大学学报,2016,39(4):632-639.[doi:10.7685/jnau.201509030]
 JIN Wenwen,WANG Dedi,PAN Zengxiang,et al.c.-1760C>A variant of BMP15 gene and its association with the promoter activity in Hu sheep and Bashibai sheep[J].Journal of Nanjing Agricultural University,2016,39(4):632-639.[doi:10.7685/jnau.201509030]
点击复制

湖羊和巴什拜羊BMP15基因c.-1760C>A变异与启动子区活性的关系()
分享到:

《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
39卷
期数:
2016年4期
页码:
632-639
栏目:
出版日期:
2016-07-01

文章信息/Info

Title:
c.-1760C>A variant of BMP15 gene and its association with the promoter activity in Hu sheep and Bashibai sheep
作者:
金雯雯1 王德迪1 潘增祥1 决肯2 李齐发1
1. 南京农业大学动物科技学院, 江苏 南京 210095;
2. 新疆农业大学动物科学学院, 新疆 乌鲁木齐 830052
Author(s):
JIN Wenwen1 WANG Dedi1 PAN Zengxiang1 JUE Ken2 LI Qifa1
1. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China;
2. College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China
关键词:
BMP15基因湖羊巴什拜羊SNPs启动子区活性
Keywords:
BMP15 geneHu sheepBashibai sheepsingle nucleotide polymorphisms (SNPs)promoter activity
分类号:
S826
DOI:
10.7685/jnau.201509030
摘要:
[目的] 为了解湖羊和巴什拜羊骨形态发生蛋白15(BMP15)基因编码区和5’调控区(5’UTR)序列特征与差异,检测其SNPs(single nucleotide polymorphisms)并探索其功能。[方法] 利用克隆测序技术获得湖羊和巴什拜羊BMP15基因编码区和5’UTR序列,运用生物信息学方法分析其序列特征;采用DNA池测序法筛选湖羊和巴什拜羊BMP15基因SNPs,直接测序法检测SNPs位点多态性;采用荧光素酶报告基因系统检测启动子区活性。[结果] 湖羊和巴什拜羊BMP15基因编码区序列长度均为1182 bp,编码393个氨基酸,编码蛋白含有典型的TGF-β结构域;湖羊和巴什拜羊BMP15基因编码区序列的同源性为99.92%,发现1个SNP位点。获得1806 bp湖羊和巴什拜羊BMP15基因5’调控区序列,发现5个SNPs位点。BMP15基因c.-1760C>A位点多态性分析发现湖羊群体中均为CC基因型,巴什拜羊群体中有CC、CA和AA 3种基因型,等位基因C和A频率分别为0.7981和0.2019。启动子区活性检测显示CC型启动子区活性明显高于AA型。[结论] 湖羊和巴什拜羊BMP15基因编码区序列高度保守,c.-1760C>A变异可能影响BMP15基因启动子区活性。
Abstract:
[Objectives] To identify the characterization of 5’-untranslated region (5’-UTR) and coding region (CDS) sequence of bone morphogenetic protein15 (BMP15) gene between Hu sheep and Bashibai sheep,we analyzed the single nucleotide polymorphism (SNP) in BMP15 gene and explored its function.[Methods] The coding and 5’-untranslated sequence of BMP15 gene were obtained using cloning and sequencing technology and we used the bioinformatics methods to analyze its sequence characterization in the Hu sheep and Bashibai sheep. The SNPs of BMP15 gene were screened using DNA pool technology and were detected using sequencing method. The activity of BMP15 promoter was investigated using dul-luciferase reporter assay system.[Results] The full length coding region of BMP15 were both 1 182 bp in Hu sheep and Bashibai sheep,which encoded 393 amino acids and contained TGF-β domain. The coding regions of BMP15 in Hu sheep shared similarities of 99.92% to Bashibai sheep,in which we detected one SNP in Hu sheep and Baishibai sheep. A total of 1 806 bp sequence of 5’-UTR was obtained,where we identified five SNPs in Hu sheep and Baishibai sheep. There was one genotype (CC) and three genotypes (CC,CA,AA)(c.-1760C>A) in the 5’-UTR of BMP15 gene in Hu sheep and Bashibai sheep,respectively. Further analysis showed that the frequency of C allele and A allele were 0.798 1 and 0.201 9,respectively. The luciferase reporter assay indicated that the promoter activity of genotype CC was obviously high than genotype AA.[Conclusions] The coding region sequences of BMP15 gene are highly conservative between Hu sheep and Bashibai sheep. The SNP (c.-1760C>A) may be involved in the effect on the promoter activity of BMP15 gene.

参考文献/References:

[1] Davis G H,McEwan J C,Fennessy P F,et al. Evidence for the presence of a major gene influencing ovulation rate on the X chromosome of sheep[J]. Biology of Reproduction,1991,44(4):620-624.
[2] Paz E,Quiñones J,Bravo S,et al. Genotyping of BMPR1B,BMP15 and GDF9 genes in Chilean sheep breeds and association with prolificacy[J]. Animal Genetics,2014,46(1):98-99.
[3] Juengel J L,Hudson N L,Heath D A,et al. Growth differentiation factor 9 and bone morphogenetic protein 15 are essential for ovarian follicular development in sheep[J]. Biology of Reproduction,2002,67(6):1777-1789.
[4] Juengel J L,Quirke L D,Lun S,et al. Effects of immunizing ewes against bone morphogenetic protein 15 on their responses to exogenous gonadotrophins to induce multiple ovulations[J]. Reproduction,2011,142(4):565-572.
[5] Galloway S M,McNatty K P,Cambridge L M,et al. Mutations in an oocyte-derived growth factor gene(BMP15)cause increased ovulation rate and infertility in a dosage-sensitive manner[J]. Nature Genetics,2000,25(3):279-283.
[6] Hanrahan J P,Gregan S M,Mulsant P,et al. 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)[J]. Biology of Reproduction,2004,70(4):900-909.
[7] Bodin L,Pasquale D E,Fabre S,et al. A novel mutation in the bone morphogenetic protein 15 gene causing defective protein secretion is associated with both increased ovulation rate and sterility in Lacaune sheep[J]. Endocrinology,2007,148(1):393-400.
[8] Martinez-Royo A,Jurado J J,Smulders J P,et al. A deletion in the bone morphogenetic protein 15 gene causes sterility and increased prolificacy in Rasa Aragonesa sheep[J]. Animal Genetics,2008,39(3):294-297.
[9] Demars J,Fabre S,Sarry J,et al. Genome-wide association studies identify two novel BMP15 mutations responsible for an atypical hyperprolificacy phenotype in sheep[J]. PLoS Genetics,2013,9(4):e1003482.
[10] Zamani P,Nadri S,Saffaripour R,et al. A new mutation in exon 2 of the bone morphogenetic protein 15 gene is associated with increase in prolificacy of Mehraban and Lori sheep[J]. Tropical Animal Health and Production,2015,47(5):855-860.
[11] 吴勇聪,应诗家,闫益波,等. 绵羊BMP15基因的多态性及其与湖羊产羔量的相关性[J]. 西北农业学报,2011,20(3):3-7. Wu Y C,Ying S J,Yan Y B,et al. Genetic polymorphism of BMP15 gene and its relationship with litter size traits in Hu sheep[J]. Acta Agriculturae Boreali-Occidentalis Sinica,2011,20(3):3-7(in Chinese with English abstract).
[12] 姚勇,潘增祥,张久峰,等. 二花脸猪NR5A2基因克隆与卵巢组织转录水平分析[J]. 南京农业大学学报,2013,36(3):133-138. DOI:10.7685/j.issn.1000-2030.2013.03.023 Yao Y,Pan Z X,Zhang J F,et al. Cloning and transcriptional level of NR5A2 gene in ovaries of Erhualian pig[J]. Journal of Nanjing Agricultural University,2013,36(3):133-138(in Chinese with English abstract).
[13] Lin J Y,Pitman-Crawford J L,Bibby A H,et al. Effects of species differences on oocyte regulation of granulosa cell function[J]. Reproduction,2012,144(5):557-567.
[14] Feary E S,Juengel J L,Smith P,et al. Patterns of expression of messenger RNAs encoding GDF9,BMP15,TGFBR1,BMPR1B,and BMPR2 during follicular development and characterization of ovarian follicular populations in ewes carrying the Woodlands FecX2W mutation[J]. Biology of Reproduction,2007,77(6):990-998.
[15] Xu Y,Li E,Han Y,et al. Differential expression of mRNAs encoding BMP/Smad pathway molecules in antral follicles of high- and low-fecundity Hu sheep[J]. Animal Reproduction Science,2010,120(1/2/3/4):47-55.
[16] Crawford J L,Heath D A,Reader K L,et al. Oocytes in sheep homozygous for a mutation in bone morphogenetic protein receptor 1B express lower mRNA levels of bone morphogenetic protein 15 but not growth differentiation factor 9[J]. Reproduction,2011,142(1):53-61.
[17] 祁艳霞,张小辉,王占彬,等. 牛BMP15基因5’UTR区克隆及生物信息学分析[J]. 河南农业科学,2007(11):104-106. Qi Y X,Zhang X H,Wang Z B,et al. Clone and bioinformatic analysis of cattle BMP15 gene 5’UTR region[J]. Journal of Henan Agricultural Sciences,2007(11):104-106(in Chinese with English abstract).
[18] Hermann B P,Heckert L L. Silencing of Fshr occurs through a conserved,hypersensitive site in the first intron[J]. Molecular Endocrinology,2005,19(8):2112-2131.
[19] Mishra B,Park J Y,Wilson K,et al. X-linked lymphocyte regulated gene 5c-like(Xlr5c-like)is a novel target of progesterone action in granulosa cells of periovulatory rat ovaries[J]. Molecular and Cellular Endocrinology,2015,412:226-238.
[20] Mullen M P,Hanrahan J P,Howard D J,et al. Investigation of prolific sheep from UK and Ireland for evidence on origin of the mutations in BMP15(FecX(G),FecX(B))and GDF9(FecG(H))in Belclare and Cambridge sheep[J]. PLoS ONE,2013,8(1):e53172.
[21] 邢海燕,贾玉娇,唐克晶,等. 转录因子GATA-2对iASPP基因的转录调控研究[J]. 中国实验血液学杂志,2013,21(3):550-555. Xing H Y,Jia Y J,Tang K J,et al. Effects of transcription factor GATA-2 on transcriptive regulation of iASPP gene[J]. Journal of Experimental Hematology,2013,21(3):550-555(in Chinese with English abstract).
[22] Saito Y,Fujiwara T,Ohashi K,et al. High-Throughput siRNA screening to reveal GATA-2 upstream transcriptional mechanisms in hematopoietic cells[J]. PLoS ONE,2015,10(9):e0137079.
[23] Sher N,Yivgi-Ohana N,Orly J. Transcriptional regulation of the cholesterol side chain cleavage cytochrome P450 gene(CYP11A1)revisited:binding of GATA,cyclic adenosine 3’,5’-monophosphate response element-binding protein and activating protein(AP)-1 proteins to a distal novel cluster of cis-regulatory elements potentiates AP-2 and steroidogenic factor-1-dependent gene expression in the rodent placenta and ovary[J]. Molecular Endocrinology,2007,21(4):948-962.
[24] Yivgi-Ohana N,Sher N,Melamed-Book N,et al. Transcription of steroidogenic acute regulatory protein in the rodent ovary and placenta:alternative modes of cyclic adenosine 3’,5’-monophosphate dependent and independent regulation[J]. Endocrinology,2009,150(2):977-989.
[25] Charles M A,Saunders T L,Wood W M,et al. Pituitary-specific Gata2 knockout:effects on gonadotrope and thyrotrope function[J]. Molecular Endocrinology,2006,20(6):1366-1377.

相似文献/References:

[1]石国庆,茆达干,程瑞禾,等.湖羊和新疆细毛羊妊娠早期内分泌比较[J].南京农业大学学报,2008,31(1):146.[doi:10.7685/j.issn.1000-2030.2008.01.030]
 SHI Guo-qing,MAO Da-gan,CHENG Rui-he,et al.Comparison of endocrinology during early pregnancy of Hu-sheep and Xinjiang Fine Wool Sheep[J].Journal of Nanjing Agricultural University,2008,31(4):146.[doi:10.7685/j.issn.1000-2030.2008.01.030]
[2]谢庄,程瑞禾,韩玉刚.湖羊公羊产羔效应的统计学分析[J].南京农业大学学报,1998,21(1):77.[doi:10.7685/j.issn.1000-2030.1998.01.015]
 Xie Zhuang,Cheng Ruihe,Han Yugang.Analysis on the lambing effects of rams in Hu sheep[J].Journal of Nanjing Agricultural University,1998,21(4):77.[doi:10.7685/j.issn.1000-2030.1998.01.015]
[3]雒秋江,韩正康.高蛋白和一般蛋白日粮条件下甲醛处理大豆粉对湖羊蛋白质消化代谢的影响[J].南京农业大学学报,1986,9(03):68.[doi:10.7685/j.issn.1000-2030.1986.03.010]
 Luo Qiujiang,Han Zhengkang.EFFECTS OF FORMALDEHYDE-TREATED GROUND SOYBEAN ON DIGESTION AND METABOLISM OF PROTEIN IN HUZHOU SHEEP FED ON TWO DIETS WITH DIFFERENT PROTEIN LEVELS[J].Journal of Nanjing Agricultural University,1986,9(4):68.[doi:10.7685/j.issn.1000-2030.1986.03.010]
[4]岳根华,谢成侠,程瑞禾,等.湖羊及考力代公羔生殖系统发育与精子发生的研究[J].南京农业大学学报,1993,16(03):69.[doi:10.7685/j.issn.1000-2030.1993.03.014]
 Yue Genhua Xie Chengxia Cheng Ruihe Zhang Quanfu Xu Subiao.STUDY ON THE DEVELOPMENT OF REPRODUCTIVE SYSTEM AND SPERMATOGENESIS IN HU SHEEP AND CORRIEDLE MALE LAMBS[J].Journal of Nanjing Agricultural University,1993,16(4):69.[doi:10.7685/j.issn.1000-2030.1993.03.014]
[5]汤艾非,韩正康.湖羊奶及人工诱发的乳腺分泌物中某些生化物质的动态变化[J].南京农业大学学报,1993,16(00):25.[doi:10.7685/j.issn.1000-2030.1993.00.006]
 Tang Aifei Han Zhengkang.OBSERVATION ON THE DYNAMICAL CHANGES OF BIOCHEMICAL COMPOSITION OF MILK AND HORMONE-INDUCED MAMMARY SECRETIONS IN HU SHEEP[J].Journal of Nanjing Agricultural University,1993,16(4):25.[doi:10.7685/j.issn.1000-2030.1993.00.006]
[6]沈延法,韩正康.湖羊复胃大小肠消化代谢及吸收的研究[J].南京农业大学学报,1993,16(00):46.[doi:10.7685/j.issn.1000-2030.1993.00.010]
 Shen Yanfa Han Zhengkang.STUDIES ON DIGESTION,METABOLISM AND ABSORPTION IN STOMACH,SMALL INTESTINE AND LARGE INTESTINE OF HU SHEEP[J].Journal of Nanjing Agricultural University,1993,16(4):46.[doi:10.7685/j.issn.1000-2030.1993.00.010]
[7]刘晳洁,韩正康.灌注乙酸研究湖羊瘤胃与盲肠的相互联系[J].南京农业大学学报,1993,16(00):50.[doi:10.7685/j.issn.1000-2030.1993.00.011]
 Liu Xijie Han Zhengkang.STUDIFS ON THE COORDINATION BETWEEN RUMEN AND CECUM BY INFUSING ACETIC ACID IN HU SHEEP[J].Journal of Nanjing Agricultural University,1993,16(4):50.[doi:10.7685/j.issn.1000-2030.1993.00.011]
[8]刘宗柱,沈延法.去势及灌注半胱胺对湖羊消化代谢的影响[J].南京农业大学学报,1996,19(2):54.[doi:10.7685/j.issn.1000-2030.1996.02.012]
 Liu Zongzhu,Shen Yanfa,Han Zhengkang.EFFECTS OF CASTRATION AND CYSTEAMINE ON DIGESTION AND METABOLISM OF HU SHEEP[J].Journal of Nanjing Agricultural University,1996,19(4):54.[doi:10.7685/j.issn.1000-2030.1996.02.012]
[9]刘吉英,郝称莉,李伟华,等.湖羊肌肉MC4R基因表达水平与肌内脂肪含量的相关性分析[J].南京农业大学学报,2011,34(6):143.[doi:10.7685/j.issn.1000-2030.2011.06.026]
 LIU Ji-ying,HAO Cheng-li,LI Wei-hua,et al.Association of the MC4R gene expression with intramuscular fat content in Hu sheep muscles[J].Journal of Nanjing Agricultural University,2011,34(4):143.[doi:10.7685/j.issn.1000-2030.2011.06.026]
[10]马铁伟,王强,王锋,等.营养水平对湖羊生长性能、血清生化指标、屠宰性能和肉品质的影响[J].南京农业大学学报,2016,39(6):1003.[doi:10.7685/jnau.201511011]
 MA Tiewei,WANG Qiang,WANG Feng,et al.Effects of different nutrition levels on growth performance,serum indexes,slaughter performance and meat traits of Hu lamb[J].Journal of Nanjing Agricultural University,2016,39(4):1003.[doi:10.7685/jnau.201511011]

备注/Memo

备注/Memo:
收稿日期:2015-09-25。
基金项目:江苏省自然科学基金项目(BK20141369);江苏省农业科技自主创新资金项目(CX(15)1007)
作者简介:金雯雯,硕士研究生。
通信作者:李齐发,教授,主要从事动物分子遗传与育种研究,E-mail:liqifa@njau.edu.cn。
更新日期/Last Update: 1900-01-01