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锌指蛋白在调节植物防卫基因表达和抗性反应上起关键作用。目前,对大豆中C3HC4型RING锌指蛋白基因的研究不多。本研究利用核蛋白筛选系统(NTT)筛选大豆(铁丰8号)干旱处理5h的cDNA文库,获得一个RING锌指蛋白基因。该基因全长927bp,编码308个氨基酸,含有C3HC4-type RING锌指结构域,命名为GmRZFP1。系统进化树分析显示,Gm-RZFP1属于C3HC4-type锌指亚家族。Real-time PCR结果表明,GmRZFP1基因受干旱、高盐、高温、低温、乙烯和ABA等胁迫诱导表达,表明该蛋白涉及多种胁迫相关的信号传导途径。亚细胞定位结果表明,163hGFP-GmRZFP1融合蛋白定位于细胞核中。本研究结果有助于研究该类基因在大豆逆境应答反应中的作用,阐明大豆抗逆分子机制。
Zinc finger proteins play a key role in regulating plant defense gene expression and resistance. At present, little research has been done on the gene of C3HC4 RING zinc finger protein in soybean. In this study, we screened the cDNA library of soybean (Tiefeng No.8) treated with drought for 5h by using the nuclear protein screening system (NTT) to obtain a RING zinc finger protein gene. The gene is 927 bp long and encodes 308 amino acids. It contains the C3HC4-type RING zinc finger domain and is named GmRZFP1. Phylogenetic tree analysis showed that Gm-RZFP1 belonged to the C3HC4-type zinc finger subfamily. Real-time PCR results showed that the GmRZFP1 gene was induced by drought, high salt, high temperature, low temperature, ethylene and ABA stress, indicating that the protein involved in a variety of stress-related signaling pathways. Subcellular localization results show that the 163hGFP-GmRZFP1 fusion protein is localized in the nucleus. The results of this study are helpful to study the role of these genes in soybean response to stress and elucidate the molecular mechanism of soybean resistance.