山羊肌肉差异表达miRNA表达规律分析
研究在山羊生长发育过程中在肌肉中高表达的miRNA，其miRNA可能参与肌肉细胞增殖与分化相关的一些靶基因的表达，为进一步研究该高表达量的miRNA参与山羊生长发育调控功能提供基础资料，从而影响山羊的生长发育。
毕业论文关键词：徐淮山羊、肌肉、miRNA、表达分析
Goats muscle differentially expressed micrornas expression pattern
Abstract:This study is to analyze our previous experiment from five known micrornas selection: miR - 323 - a - 3 p, miR - 541-3 p, let - 7 e, miR - 424-5 p, miR - 432 and five new predicted of micrornas: NovelmiR - 26, NovelmiR-65,NovelmiR-30,NovelmiR -70,NovelmiR-84, with Xu Huai goats different development period (90 days fetal sheep and 6 months of age young sheep),Their longest back muscle, heart, liver, spleen, lung, kidney, small intestine and seven tissue samples for material, Using the qRT-PCR method to detect the miRNA expression in the goat organizations, on the basis of the resulting expression spectrum analysis of the 10 kinds of micRNA expression quantity in each group. After a graph to compare the results show that in fetal sheep the Let - 7 e, miR - 432, NovelmiR - 30, NovelmiR – 26, this several micrornas quantity is higher than that of other organizations, and in the young sheep that their expression quantity about the Let - 7 e, miR - 29 a, miR - 432, and NovelmiR – 30are higher, which can be seen that the Let - 7 e, miR - 432, NovelmiR - 30 in fetal sheep and young sheep muscle are in higher quantity.
Research in goats in the process of growth and development have high expression of micrornas in muscle, the miRNA may participate in the muscle cell proliferation and differentiation of related some of the expression of target genes, in order to further study ,the high expression of micrornas are involved in goat growth regulation function providing the basic data, so as to affect the growth and development of goats.
Keywords: Xu Huai goats, muscle, micrornas, expression analysis
目录
引言 1
1材料与方法 2
1.1 实验动物及组织样品的采集 2
1.1.1 试验动物样本来源 2
1.1.2 所需主要实验试剂 2
1.2 试验方法 2
1.2.1总RNA的提取及质量检测 2
1.2.2总RNA反转录合成cDNA 2
1.2.3荧光实时定量PCR 3
2结果与分析 4
2.1RNA提取结果检测 5
2.2 miRNA在不同组织中的相对表达量分析 5
2.2.1miRNA表达谱 5
2.2.2表达量结果分析 8
3讨论 9
4结论 10
致谢 11
参考文献 11
引言
miRNA是一类内源性的长约22nt非编码的小分子RNA，可以通过与mRNA的3′非翻译区（3′-UTR）的碱基互补配对，引起靶mRNA降解、稳定性下降或者抑制其翻译，然后对基因转录后的表达进行调控，从而参与动物的生殖、生长、发育等多种生命活动[1]。研究山羊各组织中miRNA的表达谱和特征对于进一步探究它们在山羊各组织细胞增殖、分化等生长发育过程中的作用及机理都具有重要意义[2]。这些年来对miRNAs在动植物中的研究有很多，从1993年，Lee等在秀丽隐杆线虫中发现了第１个miRNA，名为lin-4，他们发现lin-4虽然可以编码RNA但不编码蛋白质，而且能阶段性调控lin-14,这个能在胚胎后期发育的基因，以此控制线虫幼虫由L1期向L2期的转化。而掀起人们对于miRNA的研究热潮是第２个被发现的miRNA分子let-7 [3]。随着高通量测序和芯片等技术的发展，越来越多的研究者把该技术应用于检测生物体不同发育阶段不同组织部位的miRNAs。目前已经开展了不少对miRNAs表达谱的研究，比如猪、鸡、牛、绵羊、山羊的，也发现并鉴定了一些物种新的miRNAs，为解析miRNAS的功能及其靶基因研究奠定了基础[4]。
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