TcLr35小麦中病程相关蛋白1基因的克隆及分析

Cloning and Characterization of Pathogenesis-related Protein 1 Gene from TcLr35 Wheat

根据已发表的植物病程相关蛋白1基因设计引物,利用RT-PCR技术,从被小麦叶锈菌诱导的小麦抗叶锈病基因近等基因系材料TcLr35中获得一个病程相关蛋白1基因cDNA片段,长度为489bp,3′端包含21个poly(A),暂命名为PR12。利用5′RACE技术获得了818bp的PR12全长,该基因包含495bp的开放读码框,147bp的5′非翻译区(non translated region,NTR),155bp的3′非翻译区和21bp的多聚腺苷酸尾。编码164个通读的蛋白质氨基酸序列,基因产物具有植物防御体系中病程相关蛋白SCP保守结构域,与GenBank中多个植物病程相关蛋白1基因具有较高的同源性。Southern杂交显示,该基因在小麦基因组中为单拷贝。

One pair of primer was designed based on the pathogenesis-related protein 1. A wheat pathogenesis related protein 1 cDNA fragment named PR12 which is 489bp in length and contained 21-bp poly （A） tails at 3′- end in TcLr35 carrying Lr35 gene conferring resistance against wheat leaf rust induced by Puccinia triticina, was isolated and characterized using RT-PCR. A full length of PR12 was obtained through 5′RACE method. The nucleotide sequence of PR12 was 818bp in length, which included a 495bp complete open reading frame encoding 164 amino acids, a 147bp 5′ non-translated region （5 NTR）, a 155bp 3′ non-translated region （3′ UTR） and a 21-bp poly （A） tail. The deduced amino acids contained SCP conserved domain, which is related to plant defense systems. The deduced amino acid sequence showed close homology to PR-1 like proteins, which have been isolated from many plants. Southern blot analysis indicated that the wheat genome contained a single copy of PR12 gene.