| 摘要: |
| 【目的】克隆麦洼牦牛干扰素-τ（IFN-τ）基因，并检测其在麦洼牦牛不同组织和不同发情周期卵巢组织中的表达水平。【方法】采用反转录PCR（reverse transcription PCR，RT-PCR）技术克隆麦洼牦牛IFN-τ基因，测序后采用不同的生物信息学软件分析其同源性，并预测IFN-τ蛋白的结构和功能。以GAPDH基因为内参基因，采用实时荧光定量PCR（quantitative real-time PCR,RT-qPCR）检测麦洼牦牛心、肝、脾、肺、肾、胃、小肠、肌肉组织和处于不同发情周期（卵泡期、红体期、黄体期）卵巢组织中IFN-τ基因mRNA的表达水平。【结果】通过RT-PCR扩增得到长598 bp的麦洼牦牛IFN-τ基因，其开放阅读框大小为564 bp，编码氨基酸187个。IFN-τ蛋白二级结构中占比最大的为α-螺旋(50.8%)，共有15个磷酸化位点，为整体带负电荷、不稳定的亲水性蛋白，其基因序列与大额牛、黄牛有较高的同源性。组织表达谱分析结果表明，麦洼牦牛小肠、肝脏组织中IFN-τ基因mRNA表达水平显著高于其他组织（P＜0.05），卵泡期卵巢组织IFN-τ基因mRNA表达水平显著高于红体期和黄体期（P＜0.05）。【结论】克隆了麦洼牦牛IFN-τ基因，该基因在麦洼牦牛卵泡期卵巢组织中的表达水平最高，提示其可能在卵巢早期发育过程中有一定作用。 |
| 关键词: 干扰素-τ 麦洼牦牛 卵巢 组织表达 |
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| 基金项目:中央高校基本科研业务费专项基金项目（2021NYB01）；青藏高原牦牛高效安全养殖技术应用与示范国家重点研发计划项目（2018YFD0502304） |
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| Cloning of Maiwa Yak IFN-τ gene and analysis of its expression |
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GAO Shaoshuai,TANG Ziwen,LI Shan,et al
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| Abstract: |
| 【Objective】This study cloned Maiwa Yak interferon-τ (IFN-τ) gene and detected its expression levels in different tissues and ovarian tissues at different stages of estrous cycle.【Method】The IFN-τ gene of Maiwa Yak was cloned by reverse transcription PCR (RT-PCR).After sequencing,different bioinformatics software packages were used to analyze the homology of IFN-τ gene and predict the structure and function of IFN-τ protein.Using GAPDH as internal reference gene,the expression levels of IFN-τ mRNA in heart,liver,spleen,lung,kidney,stomach,small intestine and muscle tissues as well as ovarian tissues at different stages (follicular,red body and luteal phase) of estrous cycle were detected by RT-qPCR.【Result】RT-PCR amplification obtained a 598 bp Maiwa Yak IFN-τ gene with 564 bp open reading frame (ORF) and 187 amino acids.The largest proportion of IFN-τ protein secondary structure is alpha helix (50.8%),and it contained 15 phosphorylation sites and it was a negative charged and unstable hydrophilic protein.TheIFN-τ gene sequence had high homology with Bos frontalis and Bos taurus.The tissue expression profiling showed that the expression levels of IFN-τ gene mRNA in small intestine and liver of Maiwa Yak were significantly higher than those in other tissues (P<0.05).The expression level of IFN-τ gene mRNA in ovarian tissue at follicular phase was significantly higher than those in red body phase and luteal phase (P<0.05).【Conclusion】The IFN-τ gene of Maiwa Yak was cloned and its expression level in ovarian tissue was the highest at follicular phase,suggesting its functions in early ovary development. |
| Key words: IFN-τ Maiwa Yak ovary tissue expression |
