摘要本实验是运用常规的Real- time PCR 技术来研究分析湖羊成年羊中七种不同组织（背肌、腿肌、心、肝、脾、肾、肺、）miR-432表达差异性。实验操作是首先提取湖羊成年羊中七个组织总的RNA，利用miR-432的特异性引物逆转录生成cDNA，再利用Real-time PCR 检测湖羊成年羊中miR-432在七个不同组织中的表达水平。实验结果表明：miR-432 在腿肌中的表达水平最高，在肝脏、背肌、脾脏、心脏也发现有miR-432 的较高水平表达，而在肺、肾脏等组织中的表达水平较低；miR-432 在湖羊成年羊的各组织表达水平依次为腿肌（1.0000）、心（0.3719）、肝（0.3113）、脾（0.1840）、背肌（0.1048）、肾（0.0595）、肺（0.0343）。结论：使用Real-time PCR方法成功检测到miRNA-432在湖羊成年羊七个组织中的表达水平，可以推测miR-432对湖羊肌肉的发育和分化可以产生重要影响，此结果可为今后湖羊的microRNA的研究以及湖羊的遗传育种工作提供科学的理论依据和实验基础。43641

该论文有图5幅，表4个，参考文献14篇

毕业论文关键词：miR-432  差异性表达  real- time PCR

Study on Different Expression of miR-432 in Hu Sheep Issues

Abstract

 Research and analysis to adult Hu sheep Seven different tissues (back muscles, leg muscle, heart, liver, spleen, kidney, lung,) miR-432 expression differences in this experiment is the use of conventional real- time PCR technology. Experimental operation is first extracted total RNA seven organizations Hu sheep adult sheep, using specific primers for reverse transcription of miR-432 to generate cDNA, and then use Real-time PCR detection Hu sheep adult sheep miR-432 in eight different tissues the expression levels. The results showed that: miR-432 expression levels in the leg muscle of the highest in the liver, back muscles, spleen, heart also found to have higher expression levels of miR-432, and the low level of expression in the lung, kidney and other tissues ; miR-432 expression levels in various tissues of adult sheep Hu sheep were leg (1.0000), heart (0.3719), liver (0.3113), spleen (0.1840), back muscles (0.1048),lung (0.0343). Conclusion: Real-time PCR method was successfully detected miRNA-432 expression levels in the lake sheep seven adult sheep tissue, miR-432 can be speculated that Hu Sheep muscle development and differentiation can have a significant impact, this result may be in the future provide a theoretical basis and experimental basic science research microRNA lake sheep and Hu sheep genetic breeding.

Key word：miR-432  Differential Expression  real- time PCR

目  录

摘  要 I

Abstract II

图清单 IV

表清单 IV

1前言 1

2材料与方法 2

2.1材料 2

2.2实验方法 2

2.3.miR-432基因的荧光实时定量PCR[8] 4

2.4数据处理 4

3结果分析 4

3.1RNA的纯度浓度数据 4

3.2 miR-432基因的荧光定量PCR结果数据 6

3.2 miR-432基因和18s基因的扩增 7

3.3 miR-432基因表达的差异性结果 9

4讨论 10

5结论 11

参考文献 12

致谢