湖羊NR5A1基因核心启动子区突变位点筛选及其与产羔数的关联分析
毕业论文关键词：湖羊； NR5A1基因；SNPs；关联分析
Screening of Mutation Site in NR5A1 Core Promoter and Its Correlation with Lamb Rates in Hu sheep
Abstract：To find the SNPs of the core promoter region of NR5A1 gene related to the lamb rate of sheep, the core promoter region of NR5A1 gene was screened by DNA amplification combined with sequencing and then genotyped. SPSS software was used to analyze the association between different genotypes and lamb rates. The results showed that a SNP was found at the site of -309 in the promoter region of NR5A1, which constituted three genotypes of CC, CA and AA. The association analysis showed that there was a decrease trend in the lamb rates of the first, second and third conceptions of sheep with AA than those of CC and CA; what’s more, the lamb rate of the second conception of sheep with AA was lower than those of CC and CA(p<0.05). These results suggest that NR5A1 gene might have a certain effect on the number of lambs in Hu sheep, and -309 locus can be used as an effective genetic marker of the reproductive performance selection for the molecular breeding in Hu sheep.
Key words: Hu sheep；NR5A1；SNPs；correlation analysis
目 录
摘要1
关键词1
Abstract1
Key words1
引言1
1材料与方法3
1.1 试验材料 3
1.1.1 试验器材 3
1.1.2试验材料4
1.2试验方法4
1.2.1组织样的DNA提取4
1.2.2 PCR扩增4
1.2.3 SNPs筛选4
1.2.4关联分析4
1.2.5统计分析4
2结果5
2.1 NR5A1基因启动子片段扩增5
2.2 NR5A1基因位点SNPs筛选分析5
2.3 NR5A1基因位点多态特征分析6
2.4 NR5A1基因型与产羔数的关联分析7
3讨论 7
致谢8
参考文献8
图1 NR5A1基因启动子片段扩增电泳5
图2 NR5A1基因-309位点SNPs筛选6
表1 扩增NR5A1基因启动子区序列引物特征4
表2 NR5A1基因-309位点多态特征6
表3 NR5A1基因-309位点不同基因型与湖羊产羔数的关联分析7
湖羊NR5A1基因核心启动子区突变位点筛选及其与产羔数的关联分析
NR5A1（nuclear receptor subfamily 5 group A member 1）又名SF-1和Ad4BP[1]，由于其与类固醇生成有关，因此曾被命名为类固醇生成因子，是核受体NR5A家族里第一个被发现的成员，也是一种重要的转录因子。研究表明，NR5A1与哺乳动物雌性生殖活动具有密切联系[2-4]。人类NR5A1基因位于9q33，包含有7个外显子和6个内含子，属于FTZ-F1家族。FTZ-F1家族按照功能和表达模式分为NR5A1与肝脏受体激素-1基因（LRH-1）或α-胎球蛋白转录因子（FTF）。LRH-1受体主要在肝脏、胰脏和肠道等组织中表达，参与胆汁酸的代谢和胆固醇的调节。NR5A1基因主要通过下丘脑-垂体-肾上腺轴和下丘脑-垂体-性腺轴调节性腺的生成和分化。
NR5A1是一种特殊的核受体，其蛋白二级结构以β-转角和无规律卷曲为主。NR5A1基因主要由DNA结合结构域、铰链区、配体结合结构域和转录激活结构域组成，其中DNA结合结构域含有两个Cys2-Cyst锌指结构，可以作用于核受体和激素应答元件。铰链区可以连接DNA结合结构域和配体结合结构域。配体结合结构域在受体形成二聚体、通过与入核载体结合及与转录起始复合物上的转录因子和辅活化子相互作用这几个方面起重要作用。作为卵巢体细胞关键的转录调节子，NR5A1可调控类固醇急性调节蛋白、细胞色素P450胆固醇侧链裂解酶、芳香化酶、抗苗勒管激素、抑制素α亚基和黄体生成素β亚基等对卵巢类固醇生成、卵泡生长成熟有重要作用的基因的转录[2]。