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