大豆C3HC4型RING锌指蛋白基因GmRZFP1克隆与表达分析
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国家高技术研究发展计划(863计划)项目(2008AA10Z124);国家自然科学基金项目(30700508)


Isolation and Expression Pattern Assay of a C3HC4-type RING Zinc Finger Protein Gene GmRZFP1 in Glycine max (L.)
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    摘要:

    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,GmRZFP1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。

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    Zinc-finger proteins play a key role in the regulation expression of defense genes and resistance response of plant. So far, few articles reported C3HC4-type RING zinc-finger protein genes in the soybean. In this study, a RING zinc-finger protein gene was isolated from soybean (Glycine max, Tiefeng 8) cDNA library treated by drought stress for 5 h using NTT system. This gene contains 927 bp, coding 308 amino acids, and has a C3HC4-type RING finger domain and named GmRZFP1. Phylogenetic analysis showed that GmRZFP1 belong to C3HC4-type Zinc-finger subfamily. Real-time PCR results showed that the expression of GmRZFP1 gene was induced by drought, high salt, high temperature, low temperature, ethylene and ABA stresses, suggesting that GmRZFP1 might involve in some stresses related signal pathways. Subcellular localization assay indicated that the 163GFP-GmRZFP1 fusion protein was located in the nucleus of plant. The results would be useful in the study of functions of this gene family in stress responses of soybean, and clarify molecular mechanisms of stress resistance in soybean.

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吴学闯,曹新有,陈明,等.大豆C3HC4型RING锌指蛋白基因GmRZFP1克隆与表达分析[J].植物遗传资源学报,2010,11(3):343-348.

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