绵羊胃肠道组织中miR-1197的表达差异性研究
毕业论文关键词：miR-1197；绵羊肠胃组织；差异性表达；real-time PCR
The study of the different expression of miR-1197 in different Ovine’ issues
Abstract: To apply real-time PCR method to measure the different expression of miR-1197 in ten kinds of intestine and stomach issues of ovine and explore its significance. first,extract the total RNAs from the ten kinds of intestine and stomach issues of ovine ,and then reverse them into cDNAs respectively with the specific and random primers. The expression of miR-1197 in ten issues would be detected by real-time PCR.Being detected by real-time PCR, The expression of miR-1197 is highest in the issue of Jejunum and lowest in the issue of Rectal and the difference is 610 times.Besides,miR-1197 also has high level expression in ovine’s other issues,such as Omasum, Duodenum ,Cecum, Ileum.So miR-1197 is enriched in Ileum, Omasum,Cecum,Duodenum. Conjecture is that miR-1197 is of great significance in the development, differentiation ,functional maintenance and the occurrence of related diseases in these organs.Another conjecture is that the function of miR-1197 have something in common with Jejunum whose function is digestion and assimilation and its function is to promote the secretion of proteins and enzymes whose functions are the similar.
Key words: miR-1197; ten issues; different expressions; real-time PCR
目 录
引言 1
1 材料与方法 2
1.1 材料 2
1.1.1 标本 2
1.1.2 仪器及试剂 2
1.2 方法 3
1.2.1 RNA提取 3
1.2.2 检测提取出的RNA的浓度 3
1.2.3 总RNA逆转录成cDNA 4
1.2.4 PCR扩增、琼脂糖凝胶电泳 4
1.2.5 实时荧光定量PCR 4
1.3 统计方法 6
2 结果与分析 6
2.1 浓度测定与反转录 6
2.2 胃肠组织中miR-1197的差异性表达 8
3 讨论 13
致谢： 14
参考文献： 15
引言
MiRNA是一种短的由高等真核生物基因组编码的单链非编码RNA，近年来发现它是存在于多种真核细胞及病毒中的一类主要存在于内源性染色体上，长度一般在21～25nt的短序列。MiRNA在进化上具有高度的保守性，miRNA通过和靶基因mRNA碱基配对引导沉默复合体（RISC）降解mRNA或阻碍其翻译，从而能够对遗传物质的载体DNA片段在转录后的水平上进行的表达调控，提高或降低其生成mRNA的水平[1]，而且它还能够参与细胞的发育、增殖调控和分化并发挥着重要的作用[2]。
MiRNA在物种进化过程中表现出高度的保守性，其只在特定的组织和发育阶段进行表达，所表现出的高度的保守性、时序性和组织特异性就决定了它在细胞和组织的功能特异性，在细胞生长和发育过程的调节过程的多种作用。尽管在10年前之前才发表第一篇有关miRNA 的论文，但是直到最近miRNA的普遍性和重要性才逐渐被人们所认识。
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