GPR54基因C-816T和T-754C的多态性与牛性成熟关联性研究

发布时间：2018-10-23 07:09

【摘要】：GPR54基因是牛性成熟状的重要候选基因之一,本文选择42头安徽本地黄牛、44头西门塔尔牛及其杂交育种群116头作为试验群体,分析牛GPR54基因5’调控区的SNPs以及不同基因型启动子的启动效率和牛性成熟状之间的关联性。试验提取牛血液基因组后,通过测序鉴别SNP位点,双荧光素酶报告基因验证其SNP位点对启动子效率的影响。结果表明:1.GPR54基因的启动子区GPR973序列存在两个SNPs位点,分别是距离CDS起点上游第816位和754位:C-816T和T-754C,西门塔尔牛和安徽地方黄牛分别只有1种基因型,西门塔尔牛的基因型全部为CCTT,安徽地方黄牛的基因型全部为TTCC,分析其单倍型结果显示连锁不平衡。2.C-816T突变位点附近存在TCF-1和AP-3(2)转录因子结合位点,T-754C附近及其本身存在TFIID、NF-1、CAC-binding_pro、TCF-1转录因子以及F位点等。从C-816T到T-754C形成一个以TCF-1开头结尾的复合转录因子结合区域,其中还包含一个TFIID结合的G/TATAAA盒。经过预测,本目的片段含有一个从-878bp至-638bp的启动子区域。3.通过双荧光素酶报告基因验证了西门塔尔牛和安徽地方黄牛不同GPR973基因型的启动子效率,报告基因验证两种单倍型的启动子存在明显差异,其中-816CT-754启动子的效率要比-816TC-754提高34.31%(p0.01)。4.通过对不同品种牛GPR54进行实时荧光定量PCR反应,结果显示与双报告基因检测结果一致。C-816CT-754T牛和T-816TC-754C牛的表达差异明显(p0.05),其中C-816CT-754T牛是T-816TC-754C牛的2.6倍。5.通过对不同品种牛进行关联分析,基因型为816CC754TT牛的初情期与基因型为816TT754CC牛的初情期相比提前了1.28月,且差异极显著(p0.01)。
[Abstract]:GPR54 gene is one of the important candidate genes for sexual maturation of cattle. 42 native Anhui cattle, 44 Simmental cattle and their cross breeding population were selected as experimental populations. The relationship between the SNPs of the 5 'regulatory region of bovine GPR54 gene, the promoter efficiency of different genotypes and the sexual maturity of cattle was analyzed. After the bovine blood genome was extracted, the SNP sites were identified by sequencing, and the effect of the SNP locus on the promoter efficiency was verified by double luciferase reporter gene. The results showed that there were two SNPs loci in the promoter region of 1.GPR54 gene, 816 and 754 from the upstream of CDS: C-816T and T-754C. Simmental and Anhui yellow cattle had only one genotype, respectively. The genotypes of Simmental cattle were all CCTT,. All the genotypes of Anhui native cattle were TTCC,. The results of linkage disequilibrium showed that there were TCF-1 and AP-3 (2) transcription factor binding sites near the 2.C-816T mutation site, and T-754C region and its own existence. In TFIID,NF-1,CAC-binding_pro,TCF-1 transcription factors and F loci, and so on. From C-816T to T-754C, a complex transcription factor binding region ending with TCF-1 is formed, which also contains a TFIID binding G/TATAAA box. It is predicted that this fragment contains a promoter region from-878bp to-638bp. The promoter efficiency of different GPR973 genotypes in Simmental cattle and Anhui native cattle was verified by double luciferase reporter gene. The promoter efficiency of 816CT-754 promoter was increased by 34.31% (p0.01) compared with that of 816TC-754. The results of real-time fluorescence quantitative PCR reaction on GPR54 of different breeds of cattle showed that the results were consistent with the results of double reporter gene detection. The difference between C-816CT-754T cattle and T-816TC-754C cattle was significant (p0.05), and C-816CT-754T cattle were 2.6 times as much as T-816TC-754C cattle. According to the correlation analysis of different breeds of cattle, the initial oestrous period of 816CC754TT cattle was 1.28 months earlier than that of 816TT754CC cattle, and the difference was very significant (p0.01).
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823

【相似文献】

中国期刊全文数据库 前10条

1 雷秋霞,李善立,杨艳;鸡性成熟研究进展[J];家禽科学;2005年11期

2 韩威;李慧芳;朱云芬;宋迟;陶志云;;动物性成熟启动基因网络和神经内分泌调节研究进展[J];家畜生态学报;2012年06期

3 ;雄小鼠气味影响雌鼠性成熟的研究[J];动物学报;1976年02期

4 小幡正树;金贵发;王葆茹;;幼年豚鼠与雄性豚鼠同居对性成熟的影响[J];上海畜牧兽医通讯;1982年01期

5 刘权 ,田纯浩 ,张素芬;影响鱼类性成熟的环境因素分析[J];农村养殖技术;2002年03期

6 高健;王德芬;;鱼类性成熟的免疫控制[J];水利渔业;1992年02期

7 宁中华，李吉祥，谢德润，张维汉;“农大褐”最佳性成熟体重的研究[J];中国畜牧杂志;1994年03期

8 韩威;李慧芳;朱云芬;宋迟;陶志云;;动物性成熟启动的调控机理[J];家畜生态学报;2012年05期

9 吴鹤洲;浙江近海大黄鱼性成熟与生长的关系[J];海洋与湖沼;1965年03期

10 陈万青;;黄海湰鱼性成熟特性的初步观察[J];海洋湖沼通报;1982年01期

中国重要会议论文全文数据库 前9条

1 王洪进;王春阳;侯秋玲;刘海港;窦文文;郭慧君;;性成熟前小鼠卵泡卵子供体发育潜能低下与雌激素受体分布表达相关性研究[A];全国动物生理生化第十一次学术交流会论文摘要汇编[C];2010年

2 芦燕;王安;;饲粮核黄素和低温对育成蛋鸭抗氧化状况和性成熟的影响[A];中国畜牧兽医学会动物营养学分会第十次学术研讨会论文集[C];2008年

3 孙彦启;宋喜书;赵娟;;獭兔繁殖技术[A];“创新驱动、生态养殖、转型发展”论文集[C];2014年

4 常建伟;关宏;阎凤祥;郑立;张硕;单德海;徐慧娟;朝格巴达日胡;安晓荣;侯健;;性成熟前羔山羊卵泡的激素诱导及生殖激素水平测定[A];中国畜牧兽医学会养羊学分会2014年全国养羊生产与学术研讨会议论文集[C];2014年

5 倪迎冬;赵茹茜;;生殖轴激素对畜禽性成熟的调节[A];全国动物生理生化第九次学术交流会论文摘要汇编[C];2006年

6 韩威;李慧芳;朱云芬;宋迟;徐文娟;;鸡类KISS-1基因克隆及EST序列表达验证分析[A];第三届（2012）中国黄羽肉鸡行业发展大会会刊[C];2012年

7 单春华;粱士儒;王博;安铁洙;;性成熟前不同日龄小鼠卵泡的组织学观察[A];中国畜牧兽医学会动物解剖学及组织胚胎学分会第十四次学术研讨会论文集[C];2006年

8 ，

本文编号：2288440