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2025年7月30日 星期三
  2017, Vol. 25 Issue (4): 628-638    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
鸽PPARγ基因的克隆、序列分析及组织表达特征
田勇1,李国勤1,陶争荣1,沈军达1,陈黎1,曾涛2,钟声亮3,卢立志1
1. 浙江省农业科学院
2. 浙江省农业科学院 畜牧兽医研究所
3.
Molecular Cloning, Sequence Analysis and Tissue Distribution of PPARγ Gene in Pigeon (Columba livia)
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摘要 过氧化物酶体增殖物激活受体γ (peroxisome proliferator activated receptor γ, 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(GenBank 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 kD,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。
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田勇
李国勤
陶争荣
沈军达
陈黎
曾涛
钟声亮
卢立志
关键词 过氧化物酶体增殖物激活受体γ (PPARγ)克隆序列分析组织表达    
Abstract:Abstract Peroxisome proliferator-activated receptor γ (PPARγ) is a ligand-activated nuclear transcriptional factor, which plays an important role in regulating lipid metabolism. This study was carried out to clone the full-length cDNA of pigeon (Columba livia) PPARγ gene, analyze the protein structures, physical, chemical properties and their functional domain structures which encoded by PPARγ gene, and reveal its tissue expression pattern. In our experiment, rapid-amplification cDNA ends (RACE) was used to obtain pigeon PPARγ cDNA. The structure and function of PPARγ gene was analyzed by bioinformatics methods. Real-time PCR was performed to detect PPARγ mRNA expression levels in various tissues. The results showed that the full-length of pigeon PPARγ (GenBank No. KU166859.2) cDNA was of 1 846 bp including a 1 428 bp ORF, 63 bp 5'UTR and 355 bp 3'UTR. The ORF encoded a polypeptide of 475 amino acids, with a calculated relative molecular mass of 54.438 79 kD, pI of 6.19 and an average of hydropathicity -0.287. The predicted protein of pigeon PPARγ was composed of 7 661 atoms with the formula of C2436H3840N644O714S27. Structure prediction showed that there were 2 zinc finger structures and a ligand binding domain in the pigeon PPARγ protein. The deduced amino acid sequence of pigeon PPARγ shared 97%, 97%, 96%, 96%, 96%, 96%, 96%, 92%, 91%, 90% and 64% identities with those of Cuculus canorus, Anas platyrhynchos, Melopsittacus undulates, Taeniopygia guttata, Meleagris gallopavo, Gallus gallus, Coturnix japonica, Sus scrofa, Homo sapien, Mus musculus and Danio rerio, repectively. Phylogenetic relationship analysis indicated that all birds and mammals each formed a large branch, and zebrafish formed another independent branch. Columba livia was closely related with Cuculus canorus and Anas platyrhynchos, which was similar to the results of the biological classification. Real-time PCR analysis revealed that the mRNA level of pigeon PPARγ was highest in lung and abundant in kidney, liver, spleen and heart, while relatively low in muscle. The results showed the full-length cDNA sequence, molecular characteristics and tissue-specific distribution of pigeon PPARγ, which suggested that the PPARγ gene might have various effects on different tissues of pigeion, and also provide theoretical foundation for the further study of the structure and function of PPARγ.
Key wordsPigeon    Peroxisome proliferator-activated receptor γ (PPARγ)    Cloning    Sequence analysis    Tissue expression
收稿日期: 2016-08-13      出版日期: 2017-03-31
基金资助:浙江省农业(畜禽)新品种选育重大科技专项;国家科技部富民强县专项子课题;浙江省农科院鸽产业团队科技特派员项目
通讯作者: 卢立志     E-mail: llz@163.com;lulizhibox@163.com
引用本文:   
田勇 李国勤 陶争荣 沈军达 陈黎 曾涛 钟声亮 卢立志. 鸽PPARγ基因的克隆、序列分析及组织表达特征[J]. , 2017, 25(4): 628-638.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2017/V25/I4/628
 
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