小白菜BcUBCE2基因的克隆及表达分析

采用cDNA-AFLP和RACE技术从小白菜中克隆得到泛素结合酶E2基因(ubiquitin conjugating enzyme E2),命名为BcUBCE2。序列分析表明,BcUBCE2基因cDNA全长830bp,包含1个456bp的开放阅读框,编码152个氨基酸。结构分析发现,该序列包含一个泛素结合酶E2活性位点和一个高度保守的半胱氨酸。进化分析显示,小白菜BcUBCE2蛋白同拟南芥E2蛋白的亲缘关系最近。qRT-PCR分析表明,BcUBCE2基因在小白菜根、茎、叶中均有表达,铜处理10d时BcUBCE2基因的表达量最高。研究认为,BcUBCE2基因可能在铜胁迫响应中发挥重要作用。

Clone and Expression Analysis of BcUBCE2 Gene from Brassica chinensis L.

The ubiquitin-conjugating enzyme E2 gene,designated as BcUBCE2,was isolated from Brassica chinensis L.by cDNA-AFLP and RACE technologies.The full length sequence of BcUBCE2 gene was 830 bp,contained an open reading frame(ORF) of 456 bp that encoded 152 amino acid residues.Structural analysis revealed that the sequence contains an E2 ubiquitin-conjugating enzyme active site and a highly conserved cysteine.Phylogenetic analysis showed that BcUBCE2 protein has the closest relative with E2 protein from Arabidopsis thaliana.qRT-PCR results showed that the existence of BcUBCE2 was detectable in root,stem and leaf.The expression level of BcUBCE2 gene reached the highest under copper stress for 10 d.This showed that the BcUBCE2 gene may play an important role in the response to copper stress.