番茄泛素蛋白基因LeEBF1和LeEBF2的克隆表达分析

通过RACE和RT-PCR方法从番茄中克隆了LeEBF1（EIN3 binding F-box protein 1）和LeEBF2（EIN3 binding F-box protein 2）的全长cDNA序列，两个基因LeEBF1、LeEBF2全长分别是2 866和2 891 bp,对序列的分析表明，它们的开放阅读框分别是1 911和1 995 bp，编码区编码637和665个氨基酸残基，在氨基端含保守的F-box区域和在羧基端有14个亮氨酸重复单位，通过BLAST软件和DNAMAN分析表明这两个基因的氨基酸序列与拟南芥EBF1和EBF2有58.6％相似，同时又与其他物种的EBF蛋白的F-box区域比较有24.4％到73.2％的相近。Northern杂交指出：LeEBF1与LeEBF2在野生型和Nr的幼叶中的表达量高于成熟叶；当在果实发育期，LeEBF1与LeEBF2在青果期的表达量相比其他时期要弱。初步结果表明，LeEBF1与LeEBF2可能在番茄的生长发育中起着重要的作用。

Molecular Cloning and Characterization of LeEBF1 and LeEBF2 from Tomato

Two full-length cDNAs of LeEBF1（EIN3 binding F-box protein 1）and LeEBF2（EIN3 binding F-box protein 2）were cloned from tomato (Solanum lycopersicum) by RT-PCR and rapid amplification of cDNA ends (RACE). They were 2 866 and 2 891 bp, with open reading frame (ORF) of 1 911 bp and 1 995 bp, encoding 637 and 665 amino acids, respectively. Putative LeEBF1 and LeEBF2 proteins, each containing a well-conserved F-box motif in the amino terminus and 14 tandem leucine-rich repeats (LRRs) in the carboxyl terminus, shared 58.6% of identity with each other. Identities with other species EBF proteins within F box motif ranged from 24.4% to 73.2%. Northern blot analysis results indicated that the expression levels of LeEBF1 and LeEBF2 in young leaves were higher than those in mature leaves of wild type and Nr. During the development of fruits, LeEBF1 and LeEBF2 expressed relatively weaker at IMG stage than at other stages tested. In a conclusion, LeEBF1 and LeEBF2 may play an important role in leaf and fruit development of tomato.