鸭YAP 1基因编码区的克隆、生物信息分析及其组织表达特性

摘要

【目的】旨在克隆鸭YAP 1基因，预测其蛋白结构、功能及其组织表达特性。【方法】以北京肉鸭为材料，采用RT-PCR 技术克隆鸭YAP 1基因 CDS 区，利用多种生物信息学分析软件预测其结构与功能，并应用荧光定量 PCR检测其组织表达特性。【结果】结果表明：鸭YAP 1基因 CDS 全长1248 bp，编码415个氨基酸，氨基酸水平上与鸡相似性最高，达99．7％；氨基酸序列分析发现鸭 YAP1蛋白相对分子量为45417．5 Da，主要定位在细胞核，属水溶性蛋白，不属于分泌蛋白；预测鸭 YAP1氨基酸含有磷酸化、糖基化位点各30个；含两个 WW 结构域，且不同物种间结构域较保守；二级结构以无规则卷曲为主，三级结构呈弯曲状螺旋结构；荧光定量结果显示，YAP 1在胰脏中表达量最高，肌肉组织中最低。【结论】鸭YAP 1基因非常保守，结构、功能与哺乳动物具较高一致性。%Objective]The study was to clone the duckYAP 1 gene cDNA coding domain sequence and predict its structure,function and expression profiles.[Method]We took Peking duck for material,using RT-PCR cloned the duck YAP 1 coding domain sequence (CDS),predicted its protein structure and functions over several bioinformatics tools and analyzed its expression in 10 tissues of duck using real-time PCR.[Results]The results showed that the duckYAP 1 CDS con-sists of 1 248 nucleotides that encode 415 amino acids residues,and the amino acid homology of duck YAP 1 shares 99.7% identity with chicken.The analysis of amino acid sequence revealed that the duckYAP 1 encoded water-soluble protein and its relative molecular weight was 45 417.5 Da.Subcellular location of YAP1 was in the nucleus,and it did not belong to the secreted pro-tein.The YAP1 protein contained 30 phosphorylation sites,30 glycosylation sites and 2 WW do-mains which are relatively conserved in species.The secondary structure of YAP1 was mainly composed of random coil,while the tertiary structure of domain area showed a forniciform helix structure.The result of real-time PCR revealed that duck YAP1 expresses most in pancreas.[Conclusion]According to the results and the analyses,the structure and the function ofYAP 1 in avian have a relatively high identity with which in mammals.