TLR7 基因的克隆、鉴定及表达分析
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国家自然科学基金(31172343,31072139,31372412);公益性行业(农业)科研专项(201303033)


Molecular cloning, characterization and expression analysis of the pigeon toll-like receptor 7 gene
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    【目的】克隆鸽Toll样受体(Toll-like receptor 7, TLR7 全基因,预测其主要功能区域并分析在鸽的各种组织中的表达情况.【方法】通过 RT-PCR、RACE、相对荧光定量PCR、生物信息学软件分析方法进行研究.【结果和结论】研究发现鸽TLR7 基因cDNA全长3 516 bp,ORF全长3 175 bp,编码1 048个氨基酸.其蛋白结构主要由胞外的富含亮氨酸的结构域(LRRs)、跨膜域(TM)和胞内的 Toll/白介素 -1 受体结构域(TIR)3部分构成.鸽 TLR7 基因编码的氨基酸序列与鸿雁 Anser cygnoides、绿头鸭 Anas platyrhynchos、鸡 Gallus gallus 和麻雀 Taeniopygia guttata 的相似性均高于78%,与哺乳动物的相似性约为60%,与鱼类的相似性低于55%.鸽 TLR7 基因在小肠、脾脏、肾脏、肝脏中表达量较高,而在大脑、肺脏、气管、心脏、胰腺、肌肉、皮肤中表达量相对较低.该研究克隆了鸽 TLR7 全基因, 并预测其主要功能区域.

    Abstract:

    【Objective】 To clone and characterize the pigeon Toll like receptor 7 (TLR7) gene.【Method】RT-PCR,RACE (rapid amplification of cDNA ends),Quantitative real-time PCR and bio-informatics software was used in this study.【Result and conclusion】 The study showed that the full-length of pigeon TLR7 cDNA sequence was 3 516 bp with the longest open reading frame (ORF) of 3 175 bp encoding a peptide of 1 048 amino acids. The deduced amino acid sequence of pigeon TLR7 contained three main structural domains including an extracellular leucine rich repeats domains(LRRs),a transmembrane domain (TM) and a Toll/IL-1 receptor domain(TIR); pigeon TLR7 shared amino acid sequence similarity with the Anser cygnoides TLR7 gene, Anas platyrhynchos TLR7 gene, Gallus gallus TLR7 gene (above 78%), mammal TLR7 gene(about 60%), and fish TLR7 gene (less than 55%).Quantitative real-time PCR analysis revealed that the pigeon TLR7 mRNA widespread expression in the tested tissues was high in small intestine, spleen, kidney and low in brain,lung, trachea, heart, pancreas, muscle and skin. In this study TLR7 gene has been isolated from pigeon, cloned and its main functional domains have been predicted.

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韩翡,焦培荣,韦良孟,何静,宋亚芬,康银峰,崔进,张烁,廖明,任涛.鸽 TLR7 基因的克隆、鉴定及表达分析[J].华南农业大学学报,2014,35(3):18-23

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  • 收稿日期:2013-10-13
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  • 在线发布日期: 2014-04-15