| 牦牛MEF2A基因克隆与差异性表达分析 |
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| 引用本文： | 邓磊,柴志欣,王会,王吉坤,信金伟,姬秋梅,钟金城.牦牛MEF2A基因克隆与差异性表达分析[J].家畜生态学报,2020,41(2):14-21. |
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| 作者姓名： | 邓磊 柴志欣 王会 王吉坤 信金伟 姬秋梅 钟金城 |
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| 作者单位： | 西南民族大学青藏高原动物遗传资源保护与利用教育部重点实验室,四川成都610041;西藏自治区农牧科学院省部共建青稞和牦牛种质资源与遗传改良国家重点实验室,西藏拉萨850009 |
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| 基金项目： | 国家肉牛牦牛产业技术体系项目;创新基金 |
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| 摘 要： | MEF2A是一种由MEF2A基因编码的蛋白质,在心脏和平滑肌细胞的形态发生和肌细胞的生成过程中发挥关键作用。该研究运用RT-PCR技术并结合生物信息学分析方法,对西藏类乌齐牦牛MEF2A基因CDS区序列进行克隆及差异表达分析。结果表明:牦牛MEF2A基因CDS区全长1469 bp,共编码489个氨基酸。分子式为C 2263 H 3601 N 647 O 742 S 20,蛋白质分子量52385.58,总原子数为7273,理论等电点(pI)为6.27,消光系数为27515,不稳定指数60.68。脂溶系数和亲水性平均值分别是64.60、-0.604,该蛋白属于亲水性不稳定蛋白。蛋白质二级结构和三级结构预测结果显示,MEF2A蛋白主要由无规则卷曲、α螺旋和延长链在空间上折叠形成。牦牛MEF2A基因CDS区编码的氨基酸序列与普通牛、家猫、家犬、绵羊、小鼠、金钱豹、宽吻海豚、抹香鲸、美洲野牛、白犀、马、东北虎、猴的同源性分别为99.6%、96.1%、96.4%、99.6%、89.3%、96.8%、96.6%、96.8%、93.6%、97.2%、96.1%、96.3%、96.7%,该基因在不同物种高度保守。利用实时荧光定量PCR检测可知,MEF2A mRNA在牦牛臀肌、心脏、肝脏、肺脏中均有表达,且在臀肌中的表达优势较为显著(P<0.05)。研究结果为进一步探讨MEF2A基因对牦牛肌肉代谢调控的分子机制研究提供了理论依据。
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| 关 键 词： | 类乌齐牦牛 MEF2A基因 克隆 组织表达 REAL-TIME PCR |
Cloning and Differential Expression Analysis of MEF2A Gene in Yak |
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| DENG Lei,CHAI Zhixin,WANG Hui,WANG Jikun,XIN Jinwei,JI Qiumei,ZHONG Jincheng.Cloning and Differential Expression Analysis of MEF2A Gene in Yak[J].Acta Ecologae Animalis Domastici,2020,41(2):14-21. |
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| Authors: | DENG Lei CHAI Zhixin WANG Hui WANG Jikun XIN Jinwei JI Qiumei ZHONG Jincheng |
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| Institution: | (Key Laboratory of Qinghai-Tibet Plateau Animal Genetic Resources Protection and Utilization,the Ministry of Education,Southwest University for Nationalities,Chengdu,Sichuan 610041,China;State Key Laboratory of Barley and Yak Germplasm Resources and Genetic Improvement,Tibet Academy of Agricultural and Animal Husbandry Sciences,Lhasa,Tibet 850009,China) |
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| Abstract: | MEF2A is a protein encoded by the MEF2A gene,which plays a key role in the morphogenesis of heart and smooth muscle cells and the process of myocyte production.In this study,RT-PCR and bioinformatics analysis were used to clone and conduct differential express analysis on the sequence of the CDS region of the MEF2A gene in Tibet Lei Wuqi yak.The results showed that the CDS region of the yak MEF2A gene was 1469 bp in length and encoded a total of 489 amino acids.The molecular formula was C 2263 H 3601N 647 O 742 S 20,the molecular weight was 52385.58,the total atomic number was 7273,the theoretical isoelectric point(pI)was 6.27,the extinction coefficient was 27515,and the instability index was 60.68.The average fat solubility coefficient and hydrophilicity was 64.60 and-0.604,and the protein was a hydrophilic unstable protein.The results of protein secondary structure and tertiary structure prediction showed that MEF2A protein was mainly formed by random folding,helix and elongated chain in space.The amino acid sequence encoded by the CDS region of the yak MEF2A gene had high homology with Bos taurus,Felis catus,Canis lupus familiaris,Ovis aries,Mus musculus,Panthera pardus,Tursiops truncatus,Physrter macrocephalus,Bison bison,Ceratotherium simum,Equus caballus,Panthera tigris altaica,Macaca fascicularis,Bos grunniens,which was 99.6%,96.1%,96.4%,99.6%,89.3%,96.8%,96.6%,96.8%,93.6%,97.2%,96.1%,96.3%,96.7%,respectively.The gene was highly conserved in different species.Real-time quantitative PCR showed that MEF2A mRNA was expressed in yak gluteal muscle,heart,liver and lung,and its expression advantage in gluteal muscle was more obvious(P<0.05).Through cloning and bioinformatics and differential expression analysis,the theoretical basis of further study on the molecular mechanism of the regulation of muscle metabolism in yak is provided. |
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| Keywords: | Lei Wuqi Yak MEF2A gene clone organizational expression Real-time PCR |
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