菠萝锌指蛋白基因AcRCHY1的克隆与表达分析

 根据植物C3HC4型兼CHY型锌指蛋白的功能保守区设计简并引物, 通过RT2PCR结合RACE方法从菠萝(Ananas comosus L. Merr) 幼苗中克隆获得了一个新的菠萝锌指蛋白基因cDNA全长, 将其命名为AcRCHY1。该基因cDNA全长1 261 bp, 开放阅读框ORF为918 bp, 推测其编码一个含有306个氨基酸残基的多肽。AcRCHY1蛋白具有保守的C3HC4 (RING finger) 和CHY两个锌指结构域, 与其它植物锌指蛋白的同源性高达81%～91%。半定量RT-PCR分析表明, AcRCHY1 在菠萝中呈组成性表达, 在子房、花瓣和小花中的表达量明显高于根和叶; 低温、高盐、干旱和ABA等非生物胁迫处理后, AcRCHY1在叶片中的表达明显增强。因此, AcRCHY1蛋白可能与花器官生长发育的调控有关, 而且可能作为一个转录调控因子在菠萝响应低温、高盐和渗透胁迫过程中参与了依赖ABA的信号转导途径。

Clon ing and Expression Analysis of a Zinc Finger Protein Gene AcRCHY1 from Pineapple

Emp loying a pair of degenerate primers designed from the conserved domain of plant C3HC4 type and CHY type zinc-finger proteins, a novel zinc-finger protein gene AcRCHY1 was isolated from pineapple.(Ananas comosus) by RACE-PCR and RT-PCR techniques. The full length cDNA has 1 261 bp nucleotides with an open reading frame (ORF) of 918 bp encoding a precursor protein of 306 amino acid residues. AcRCHY1 protein sharing 81% - 91% homology with zinc-finger proteins from other plants contains two conservative zinc finger motif, C3HC4 zinc finger (RING finger) and CHY zinc finger. Semi-quantitive RT-PCR analysis indicated that AcRCHY1 was constitutively expressed in all plant organs, but stronger in ovaries, petalsand florets than in roots and leaves, and the abiotic stresses such as low temperature, high salt , drought (20% PEG6000) and ABA, could trigger a significant induction of AcRCHY1 in leaves. All these results suggest that the AcRCHY1 protein may be involved in the regulation of floral organ growth and development, and play an important role in signal transduction pathway of dependent ABA through responses to cold, high salt and osmotic stresses as a transcrip tion factor.