鸽PPARγ基因的克

发布时间：2018-04-25 18:35

  本文选题：鸽 + 过氧化物酶体增殖物激活受体γ(PPARγ)　； 参考：《农业生物技术学报》2017年04期

【摘要】：过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptorγ,PPARγ)是一种由配体激活的核转录因子,在脂类代谢调控中发挥重要作用。本研究旨在克隆鸽(Columba livia)PPARγ基因的全长cDNA序列,并分析该基因编码的蛋白质理化性质和结构特征,揭示其组织表达特征。以鸽脾脏组织总RNA为模板,采用快速扩增cDNA末端(rapid amplification cDNA ends,RACE)方法,克隆到PPARγ基因的cDNA序列,并运用生物信息学的方法对其进行分析,运用实时荧光定量PCR检测PPARγ基因在鸽心脏、肝脏、脾脏、肺、肾脏和肌肉组织中的表达情况。研究结果表明,鸽PPARγ基因(Gen Bank No.KU166859.2)cDNA全长1 846 bp,包含1 428 bp的开放阅读框,63 bp的5'端非编码区和355 bp的3'端非编码区,共编码475个氨基酸。经预测,鸽PPARγ基因编码的蛋白质由7 661个原子组成,分子式为C2436H3840N644O714S27,等电点为6.19,相对分子质量为54.438 79 k D,平均亲水性为-0.287。结构预测表明,鸽PPARγ蛋白具有2个锌指结构以及配体结合域。鸽PPARγ氨基酸序列与杜鹃(Cuculus canorus)、鸭(Anas platyrhynchos)、鹦鹉(Melopsittacus undulates)、斑胸草雀(Taeniopygia guttata)、火鸡(Meleagris gallopavo)、鸡(Gallus gallus)、鹌鹑(Coturnix japonica)、猪(Sus scrofa)、人(Homo sapien)、小鼠(Mus musculus)和斑马鱼(Danio rerio)的同源性分别为97%、97%、96%、96%、96%、96%、96%、92%、91%、90%和64%。构建的系统发生树显示,鸟类和哺乳动物各形成一个大的分支,斑马鱼形成一个独立分支,与鸽的聚类顺序较近的物种为杜鹃和鸭,该结果与这些物种在生物学上的分类是基本一致的。荧光定量PCR结果显示,PPARγ基因在肺组织中的表达量最高,较高表达于肾脏、肝脏、脾脏和心脏,而在肌肉组织中的表达量最低。本研究获得了鸽PPARγ基因的cDNA序列、分子的结构特点及组织表达特征,提示可能与其在不同组织中的功能有关,该结果为进一步研究PPARγ结构和功能提供了理论依据。
[Abstract]:Peroxisome proliferator activated receptor 纬) is a ligand activated nuclear transcription factor, which plays an important role in the regulation of lipid metabolism. The purpose of this study was to clone the full-length cDNA sequence of the livia)PPAR 纬 gene of pigeon, analyze the physicochemical properties and structural characteristics of the protein encoded by the gene, and reveal its tissue expression characteristics. Using the total RNA of pigeon spleen as template, the cDNA sequence of PPAR 纬 gene was cloned by rapid amplification of cDNA terminal amplification cDNA endsrase, and analyzed by bioinformatics. The real-time fluorescence quantitative PCR was used to detect PPAR 纬 gene in the heart of pigeon. Expression in liver, spleen, lung, kidney and muscle. The results showed that the total length of PPAR 纬 gene Gen Bank No.KU166859.2)cDNA in pigeon was 1 846 BP, which contained 1 428 BP open reading frame 63 BP 5 'uncoding region and 355 BP 3' terminal noncoding region, encoding 475 amino acids. It is predicted that the protein encoded by pigeon PPAR 纬 gene is composed of 7 661 atoms, the molecular formula is C2436H3840N644O714S27, the isoelectric point is 6.19, the relative molecular weight is 54.438 79kD, and the average hydrophilicity is -0.287. Structural prediction showed that pigeon PPAR 纬 protein had two zinc-finger structures and ligand binding domains. 楦絇PAR纬姘ㄥ熀閰稿簭鍒椾笌鏉滈箖(Cuculus canorus),楦，

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