| 摘要: |
| 【目的】克隆条锈菌诱导的小麦过敏性反应（Hypersensitive induced reaction,HIR）基因TaHIR4，分析其编码蛋白的结构、进化与功能。【方法】采用电子克隆、RT-PCR技术，从条锈菌侵染的小麦中，分离过敏性诱导基因TaHIR4，并对其进行克隆和序列分析；利用InterProScan、TMpred、SignalP 3.0、PSORT等在线分析工具，预测TaHIR4蛋白的保守结构域、跨膜结构、信号肽、细胞定位等特征；采用DNASTAR软件分别进行氨基酸序列同源性比对和进化树分析。【结果】分离到小麦过敏性诱导基因，命名为TaHIR4（867 bp，GenBank注册号为FJ028663），该基因编码1条由288个氨基酸组成的多肽；TaHIR4蛋白含HIR蛋白家族的保守SPFH（Stomatins,Prohibitin,Flotillins,HflK/C）、PHB（Prohibitin homologues）结构域，有4种典型的motifs（蛋白激酶、酪蛋白激酶Ⅱ、酪氨酸激酶磷酸化和N 豆蔻酰化位点），N末端的19个氨基酸序列内部存在跨膜信号肽结构，可能属于分泌蛋白；TaHIR4与HvHIR4蛋白的亲缘关系最近，同源性最高（98%），其次为OsHIR4，同源性为91%。【结论】分离到1个条锈菌诱导的小麦HIR基因TaHIR4，其编码蛋白可能属于分泌蛋白，在胞外行使功能。 |
| 关键词: 小麦 条锈菌 过敏性反应 电子克隆 结构域 |
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| 基金项目:国家自然科学基金项目（30930064，30800712）；农业部现代农业产业技术体系建设专项资金项目；高等学校学科创新引智计划项目（B07049）；国家“十一五”支撑计划项目（2006BAD08A05） |
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| Cloning and sequence analyses of a hypersensitive induced reaction gene TaHIR4 in wheat |
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| Abstract: |
| 【Objective】The study was conducted to clone TaHIR4,a wheat hypersensitive induced reaction gene induced by stripe rust pathogen infection and to analyze the structure,evolution and function of the encoded protein.【Method】In silico cloning and RT-PCR methods were taken to isolate the wheat HIR gene in Xingzi9104 infected by Puccinia striiformis f.sp.tritici (PST).Characteristics of the conserved domain,transmembrane structure,signal peptide,and localization of the encoded TaHIR4 protein were determined using bioinformatics analyses including InterProScan,TMpred,SignalP 3.0,and PSORT,respectively.The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR software package.【Result】The wheat HIR gene,designated as TaHIR4 (867 bp),was isolated,and deposited in GenBank with accession number of FJ028663.TaHIR4 was deduced to encode a protein of 288 amino acids,which contained two typical conserved SPFH (Stomatins,Prohibitin,Flotillins,HflK/C),PHB (Prohibitin homologues) domains.TMpred and SignalP analyses revealed the presence of putative signal peptide within 19 residues at the N-terminus of the TaHIR4 protein,suggesting the putatively secretory character.TaHIR4 protein also contained four typical kinds of motifs (N-myristoylation sites,protein kinase C,tyrosine kinase and casein kinaseⅡ phosphorylation sites).The analyses of multiple alignment and phylogenetic tree demonstrated that TaHIR4 was most closely related to barley HvHIR4 protein with 98% high similarity,followed by rice OsHIR4 protein with 91% high similarity.【Conclusion】The TaHIR4 gene was isolated from wheat infected by PST and characterized using a series of bioinformatics analyses,whose encoding protein may be secretory and function extracellularly. |
| Key words: wheat stripe rust fungus hypersensitive response in silico cloning domain |
