摘要本实验使用实时定量PCR技术来研究分析ACTC1基因在徐淮山羊胎羊八种不同组织（心、背肌、腿肌、肝、脾、肾、肺、肠）中表达差异性。实验步骤是提取胎羊八个组织总RNA，利用ACTC1基因特异性引物逆转录得到cDNA，最后用Real-time PCR 检测ACTC1基因在八个不同组织中的表达水平。实验结果表明：ACTC1基因在八个组织中都有表达，但是表达水平各不相同。在心肌组织中的表达水平最高，在背肌、腿肌也发现有少量的表达，但在肝、脾、肾、肺、肠等组织中几乎没有表达；ACTC1基因在徐淮山羊胎羊的各组织表达水平依次为心肌（1.00000）、腿肌（0.04723）、背肌（0.01445）、脾(0.00002)、肠(0.00007)、肾(0.00028)、肺(0.00002)、肝(0.00194)。采用Real-time PCR技术成功检测到ACTC1基因在山羊胎羊肌肉组织中表达量较高，可以推测ACTC1基因对于肌肉的增殖分化、功能维持可能产生要紧的影响。此结果可为以后ACTC1基因的研究提供实验基础和理论依据。50720

毕业论文关键词：徐淮山羊  ACTC1基因  PCR  差异分析

The study on different expression of ACTC1 gene in Xuhuai fetal goat organs

Abstract

We are using the Real-time PCR technology to research ACTC1 gene express differences in eight different organizations (heart, back muscle, leg  muscle, liver, spleen, kidney, lung, intestine) in Xuhuai fetal goat in this experiment. Experimental operation is picking up fetal goat’s eight organizations total RNA, using the ACTC1 gene specific primers to reverse transcription into cDNA. At last, using Real - time PCR to test ACTC1 gene expression level in eight different organizations.

 The experimental results show that ACTC1 gene express successfully expressed in eight tissues , but the quantities of expression are different. The highest level of expression is in the heart. Back muscle and leg muscle also found that the low level expression of ACTC1 gene, and almost no expression in the liver, spleen, kidney, lung, intestine. The ACTC1 gene expression levels in eight organizations are: heart（1.00000）、leg  muscle（0.04723）、back muscle（0.01445）、spleen (0.00002)、intestine (0.00007)、kidney (0.00028)、lung (0.00002)、liver (0.00194). The use of Real-time PCR detected the ACTC1 gene express higher in muscle organization successfully. It is supposed that ACTC1 gene play a important role in muscle’s proliferation differentiation、function to maintain. The results can provide experimental basis and theoretical basis for ACTC1 gene’s future research.

Key words: Xuhuai goat  ACTC1 gene  PCR  variation analysis

目  录

摘要 I

Abstrcat II

目录 III

图清单、表清单 IV

1引言 1

2 步骤、材料和方法 1

2.1步骤 1

2.2 材料 1

2.2.1样本 1

2.2.2器材设备 2

2.2.3 试剂 2

2.3方法 2

2.3.1提取各组织的总RNA 2

2.3.2检测提取出的RNA的浓度 3

2.3.3 RNA逆转录成cDNA 3

2.3.4 PCR扩增、琼脂糖凝胶电泳 3

2.3.5荧光实时定量PCR 4

3数据统计分析 5