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棉花学报  2011 

海岛棉转录因子EREB5基因的克隆及特征研究

DOI: 1002-7807(2011)03-0205-07, PP. 205-211

Keywords: 海岛棉,ERF,黄萎病,EREB5,基因克隆

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Abstract:

近年来,ERF(Ethylene-responsiveelementbindingfactor)家族转录因子已成为植物抗逆、抗病的分子机制和作物分子育种研究的热点。本研究以电子克隆、同源扩增和RACE(Rapid-amplificationofcDNAends)技术相结合的方法从海岛棉中分离出了一条新的ERF族转录因子基因,命名为EREB5。该基因包含一个573bp长的开放阅读框,预期编码190aa长的蛋白质分子。序列分析表明该蛋白除含有一个ERF保守域外,还含有一段核定位信号序列、一段富丝氨酸的激活域、多种磷酸化位点和一个土豆抑制子Ⅰ型家族基序等。构建系统发育树分析表明该因子属于ERF亚家族B3亚组。瞬时表达实验证明该因子定位于细胞核内,同时,凝胶阻滞实验结果说明EREB5蛋白和GCC盒具有较强的结合能力。再者,荧光定量PCR结果表明乙烯和黄萎病菌处理可以诱导该基因的表达。这些结果暗示EREB5蛋白很可能在黄萎病抗性机制中扮演者重要角色。

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