| 摘要: |
| 【目的】建立一种牦牛TLR1基因表达量的荧光定量PCR检测方法，并分析TLR1基因在牦牛不同组织中的表达差异。【方法】参考牦牛TLR1基因序列，在其保守区设计特异性引物，并以牦牛β-actin基因为内参基因建立荧光定量方法；基于该荧光定量方法分析TLR1基因在牦牛心、肝、脾、肺、肾、大肠、小肠、胃、乳腺、肌肉、卵巢11种组织中的表达水平。【结果】TLR1基因和β-actin基因扩增产物电泳结果呈现单一条带，其产物熔解曲线均为特异的单峰，表明引物具有较高的特异性。组织表达结果显示，TLR1基因在所检测的牦牛11个组织样本中均有表达，其中在肾、肝、脾、肺、卵巢、小肠中表达量较高，在胃、乳腺、心、大肠、肌肉组织中表达量较低。【结论】TLR1基因在牦牛各组织中转录水平差异较大，这可能与各组织对病原体的识别和抵抗能力有关。 |
| 关键词: 麦洼牦牛 TLR1 荧光定量PCR 组织表达谱 |
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| 基金项目:中央高校青年教师基金项目（2014NZYQN37） |
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| Expression analysis of TLR1 gene in different tissues of Maiwa yak |
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CHEN Ya-bing,LAN Dao-liang,HUANG Yong,et al
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| Abstract: |
| 【Objective】The study was designed to establish a real-time PCR assay for detection of TLR1 gene,and to further analyze the expression pattern in tissues of yaks.【Method】According to the sequence of yak TLR1 gene,the real-time PCR primers were designed,the real-time PCR was established,and the β-actin gene was chosen to normalize the expression level of TLR1 gene.The established detection method was used to analyze the expression pattern of TLR1 gene in different tissues of yak,including heart,liver,spleen,lung,kidney,large intestine,small intestine,stomach,mammary gland,muscle and ovary.【Result】The electrophoresis results showed that both TLR1 gene and β-actin gene included a signal band.The melting curve of target product showed that the product was specific to a single peak,suggesting the specificity of primers.Moreover,real time quantitative PCR assay was established and used to detect the relative expression level of TLR1 gene in different tissues,and the gene expression level in small intestine was selected as a baseline.The quantitative results showed that TLR1 gene was expressed at eleven tissues examined.TLR1 gene had high expression levels in kidney,liver,spleen,lung,ovary,small intestine and low levels in stomach,mammary gland,heart,large intestine and muscle.【Conclusion】There were differences on the transcription levels of TLR1 mRNA in different tissues,which may be associated with the ability to recognize and defend pathogens for different tissues. |
| Key words: Maiwa yak TLR1 real-time PCR expression pattern in tissues |
