毛白杨真核细胞翻译起始因子5A基因(PtoeIF5A4)的克隆与表达分析  

Cloning and Expression Analysis of the Eukaryotic Translation Initiation Factor 5A Gene(PtoeIF5A4) in Populus tomentosa

作　　者：张利姣[1,2] 张杰伟[1] 陈亚娟[1] 管阳[1] 王宏芝[1] 魏建华[1] 

机构地区：[1]北京市农林科学院北京农业生物技术研究中心,北京100097 [2]首都师范大学,北京100048

出　　处：《农业生物技术学报》2013年第8期949-956,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.30972392);国家高技术研究发展计划(863)项目(No.2011AA100201);国家重点基础研究发展规划(973)项目(No.2012CB114501)

摘　　要：真核细胞翻译起始因子5A(eIF5A)是目前已知的唯一一种含有羧腐胺赖氨酸残基的蛋白质,其在细胞的生长和增殖方面具有重要作用。但eIF5A各异形体的生物学功能尚不明晰。本研究从毛白杨(Populus tomentosa)中克隆了一个eIF5A的全长编码区序列,并根据同源性分析将其命名为PtoeIF5A4(GenBank登录号:HQ529482),其编码区与毛果杨(Populus trichocarpa)PtreIF5A4的编码区在核酸水平的序列相似性为99%。PtoeIF5A4的编码区全长483bp,共编码160个氨基酸,编码蛋白的预测分子量约为17.5kD,等电点为5.76。表达分析结果表明,该基因在根、茎及叶中均表达,在茎中的表达量最高,而且随生长时间的增长,PtoeIF5A4在茎中表达量逐渐上升,生长12个月的杨树茎次生木质部中PtoeIF5A4的表达水平是移栽1个月的4倍,且在12个月后趋于稳定,推测PtoeIF5A4在林木次生生长中起关键作用。T he eukaryotic translation initiation factor 5A （eIF5A） is the only cellular protein that contains the unique polyamine-derived amino acid, hypusine. Genetic and pharmacological evidences suggest that eIF5A is essential for cell survival and proliferation. However, the precise function of eIF5A is still unclear. In the present study, We isolated the full length PtoeIF5A4 （GenBank Accession number： HQ529482） CDS from white poplar based on bioinformation analysis. Sequence analysis showed that the sequence of PtoeIF5A4 shared 99% identity with the sequence of Populus trichocarpa. The completed ORF of PtoeIF5A4 was 483 bp and encoding 160 amino acid residues with a calculated molecular mass of 17.5 kD and a pI of 5.76. Real-time PCR results showed that PtoeIF5A4 expressed in various organs in poplar,especially in the stem. The expression of PtoeIF5A4 increased with growth time until one year. The expression of PtoeIF5A4 in 1 year P. tomentosa tree stem secondary xylem was 4 times than that of in 1 month. These results demonstrate that PtoeIF5A4 may play an important role in growth and development of trees secondary growth.

关 键 词：毛白杨 PtoeIF5A4 次生生长 

分 类 号：S511.03[农业科学—作物学]