乌苏里貉TLR8基因的分子克隆、序列分析及组织表达分析

Clone, sequence analysis and tissues expression profiles of raccoon dog Toll-like receptor 8 gene

为了解乌苏里貉TLR8基因特点及其在不同组织中的表达水平,本研究利用RT-PCR和RACE技术克隆其c DNA全长序列,应用生物学软件分析该基因及预测其编码蛋白的特征,并对该基因在乌苏里貉不同组织中的表达水平进行检测。结果显示该基因全长3 191 bp,ORF全长3 117 bp,编码1 038 aa,含有Toll-like受体家族典型的结构域特征。同源性分析显示乌苏里貉TLR8基因核苷酸序列与犬同源性最高,达99.3%,与其他食肉目哺乳动物也具有较高的同源性,在89.2%-92.9%。组织表达分析显示TLR8基因在乌苏里貉各组织中广泛表达但表达量存在差异。本研究结果为进一步研究乌苏里貉抗病毒免疫及育种相关研究奠定了基础。

In order to understand the characteristics and the expression patterns of raccoon dog TLR8 gene, the full length of TRL8 was cloned by RACE and RT-PCR techniques, the gene and the putative amino acid sequence were subsequently analyzed by bioinformatics software. Furthermore, its expression patterns in various tissues were analyzed by semi-quantitative RT-PCR. The result showed that full length of cDNA sequence was 3,191 bp, with a 3,117 bp ORF, which encoded 1,038 amino acids. Blast results revealed 99.3% similarities with canislupus familiaris, followed by 89.2% tp 92.9% similarities with other carnivores. Tissue expression analysis showed that TRL8 was widely and differently expressed in variety tissues. The study lays a foundation for further study on disease resistance while breeding of raccoon dogs.