马尾松PmTIR1基因的克隆及编码蛋白质的结构预测  

Cloning and Coding Protein Structure Prediction of TIR1 Gene from Pinus massoniana

作　　者：苏江[1] 郭天玮[1] 季孔庶[1] 

机构地区：[1]南京林业大学南方现代林业协同创新中心,南京210037

出　　处：《分子植物育种》2015年第6期1355-1362,共8页Molecular Plant Breeding

基　　金：国家"十二五"科技支撑项目(2012BAD01B02);林业公益性行业科研专项经费(201104010);江苏高校优势学科建设工程项目(PAPD);南方现代林业协同创新项目共同资助

摘　　要：TIR1(transport inhibitor response 1)基因在生长素信号转导途径中发挥重要作用。本研究采用PCR技术和RACE技术,利用Primer Premier5进行引物设计,从马尾松中克隆出TIR1基因的全长c DNA序列,命名为Pm TIR1。此c DNA全长2 446 bp,包括1 725 bp完整的ORF,能够编码含有574个氨基酸的蛋白,此蛋白相对分子质量为64 240.3 u,等电点为6.76。利用Clustal X、ANTHEPROT和DNAMAN等软件分析Pm TIR1序列及其编码的氨基酸序列,结果表明:马尾松Pm TIR1蛋白氨基酸序列与火距松TIR1蛋白同源性高达99%,Pm TIR1蛋白含有F-box结构域和多个LRR结构域;Pm TIR1蛋白的F-box结构域可能与SCFTIR1中SKP1结合,它符合F-box家族的特点。利用实时定量技术得知Pm TIR1在马尾松嫩根、嫩茎、嫩叶中都有表达,在嫩叶中表达量最高、嫩根中次之、嫩茎中最低。本研究可为生长素调控马尾松生长发育相关机理提供理论依据。TIR1(transport inhibitor response 1) plays an important role in auxin signal transduction pathway. In this study, we used primer 5 to design primers, through PCR and RACE technology, the TIR1 gene from Pinus massoniana named Pm TIR1 was cloned. Pm TIR1, having 2 446 bp in full length, contains an opening frame of1 725 bp that encodes a protein of 574 amino acids with predicted relative molecular mass of 64 240.3 u and PI of6.76. Its nucleotide sequence and the encoded amino acid sequence were analyzed with softwares of Clustal X,DNAMAN and ANTHEPROT, the results showed that :the homology of Pm TIR1 protein from Pinus massoniana and TIR1 protein from Pinus taeda was as high as 99%, Pm TIR1 contained the F-box protein domains and multiple LRR domain; F-box protein domains in Pm TIR1 may combine with SKP1 in SCFTIR1, it was consistent with the characteristics of F-box family. Using real-time quantitative techniques, we identified the amount of Pm TIR1 expression, which were the highest in leaf and the lowest in stem from P. massoniana. The study can help to discover how the auxin regulating growth and development of Pinus massoniana.

关 键 词：马尾松 生长素 TIR1(transport INHIBITOR RESPONSE 1) 序列分析 

分 类 号：S791.248[农业科学—林木遗传育种]