發(fā)布時間：2018-04-25 18:35

  本文選題：鴿 + 過氧化物酶體增殖物激活受體γ(PPARγ)��； 參考：《農(nóng)業(yè)生物技術(shù)學(xué)報》2017年04期

【摘要】：過氧化物酶體增殖物激活受體γ(peroxisome proliferator activated receptorγ,PPARγ)是一種由配體激活的核轉(zhuǎn)錄因子,在脂類代謝調(diào)控中發(fā)揮重要作用。本研究旨在克隆鴿(Columba livia)PPARγ基因的全長cDNA序列,并分析該基因編碼的蛋白質(zhì)理化性質(zhì)和結(jié)構(gòu)特征,揭示其組織表達特征。以鴿脾臟組織總RNA為模板,采用快速擴增cDNA末端(rapid amplification cDNA ends,RACE)方法,克隆到PPARγ基因的cDNA序列,并運用生物信息學(xué)的方法對其進行分析,運用實時熒光定量PCR檢測PPARγ基因在鴿心臟、肝臟、脾臟、肺、腎臟和肌肉組織中的表達情況。研究結(jié)果表明,鴿PPARγ基因(Gen Bank No.KU166859.2)cDNA全長1 846 bp,包含1 428 bp的開放閱讀框,63 bp的5'端非編碼區(qū)和355 bp的3'端非編碼區(qū),共編碼475個氨基酸。經(jīng)預(yù)測,鴿PPARγ基因編碼的蛋白質(zhì)由7 661個原子組成,分子式為C2436H3840N644O714S27,等電點為6.19,相對分子質(zhì)量為54.438 79 k D,平均親水性為-0.287。結(jié)構(gòu)預(yù)測表明,鴿PPARγ蛋白具有2個鋅指結(jié)構(gòu)以及配體結(jié)合域。鴿PPARγ氨基酸序列與杜鵑(Cuculus canorus)、鴨(Anas platyrhynchos)、鸚鵡(Melopsittacus undulates)、斑胸草雀(Taeniopygia guttata)、火雞(Meleagris gallopavo)、雞(Gallus gallus)、鵪鶉(Coturnix japonica)、豬(Sus scrofa)、人(Homo sapien)、小鼠(Mus musculus)和斑馬魚(Danio rerio)的同源性分別為97%、97%、96%、96%、96%、96%、96%、92%、91%、90%和64%。構(gòu)建的系統(tǒng)發(fā)生樹顯示,鳥類和哺乳動物各形成一個大的分支,斑馬魚形成一個獨立分支,與鴿的聚類順序較近的物種為杜鵑和鴨,該結(jié)果與這些物種在生物學(xué)上的分類是基本一致的。熒光定量PCR結(jié)果顯示,PPARγ基因在肺組織中的表達量最高,較高表達于腎臟、肝臟、脾臟和心臟,而在肌肉組織中的表達量最低。本研究獲得了鴿PPARγ基因的cDNA序列、分子的結(jié)構(gòu)特點及組織表達特征,提示可能與其在不同組織中的功能有關(guān),該結(jié)果為進一步研究PPARγ結(jié)構(gòu)和功能提供了理論依據(jù)。
[Abstract]:Peroxisome proliferator activated receptor 緯) 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 緯 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 緯 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 緯 gene in the heart of pigeon. Expression in liver, spleen, lung, kidney and muscle. The results showed that the total length of PPAR 緯 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 緯 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 緯 protein had two zinc-finger structures and ligand binding domains. 楦絇PAR緯姘ㄥ熀閰稿簭鍒椾笌鏉滈箖(Cuculus canorus),楦，

本文編號：1802479