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核农学报  2011 

烟草WRKY12基因的分离、表达与多克隆抗体的制备

, PP. 461-468

Keywords: 烟草,WRKY,分离,序列分析,原核表达

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Abstract:

在高等植物中,WRKY转录因子家族成员广泛参与植物的生长发育、代谢、生物胁迫和非生物胁迫等生理过程的调控。本研究采用同源序列克隆的方法获得烟草WRKY12基因,序列分析表明,该蛋白属于WRKY家族第2类成员,只有一个WRKY结构域,锌指结构为C-X4-C-X23-H-X1-H。构建系统发育树结果表明,它与大豆中GmWRKY6和GmWRKY12亲缘关系较近,相似性分别达50%和45%。NorthernBlot试验证明该基因表达受4℃低温胁迫诱导。将其完整编码区构建到大肠杆菌表达载体,在培养温度18℃条件下添加0.2mmol/LIPTG诱导表达融合蛋白。进一步试验获得特异性和纯度非常高的纯化蛋白和抗体。

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