柽柳ThERF1基因上游调控因子W-box互作蛋白

Proteins Interacting with Up-Stream Regulators of W-box in an ERF Gene (ThERF1) from Tamarix hispida

乙烯响应因子(ethylene response factor,ERF)参与植物逆境胁迫的应答,是在植物抗逆过程中起重要作用的一类转录因子。试验克隆了柽柳的一条ERF基因(ThERF1)的其启动子序列,其启动子中有3个W-box元件,试验通过酵母单杂交技术筛选获得了与W-box元件互作的基因ThWRKY2和ThWRKY8;进一步将ThERF1启动子中含有W-box元件、缺失W-box元件及突变W-box元件的片段构建到酵母报告载体pHIS2中,研究它们与ThWRKY2和ThWRKY8的结合能力。酵母单杂交结果表明,ThWRKY2和ThWRKY8能够与含有W-box元件的启动子片段互作,但不能与缺失及突变W-box元件的启动子片段互作。以上研究结果表明,ThWRKY2和ThWRKY8为ThERF1上游的调控基因,能够通过结合ThERF1启动子中的W-box元件来调控ThERF1基因的表达。

Ethylene response factor （ ERF ） is involved in the response to adverse environments in plants and plays important roles in stress tolerance.We cloned the promoter of an ERF gene （ThERF1） with three W-box motifs from Tamarix hispi-da.We obtained two WRKY proteins, ThWRKY2 and ThWRKY8, bind to W-box motif specifically by yeast one-hybrid as-say.ThERF1 promoter fragments without W-box motifs or with W-box and the mutated W-box motifs were cloned into pHIS2 reporter vector to analyze their bindings to ThWRKY2 and ThWRKY8 by yeast one-hybrid.ThWRKY2 and ThWRKY8 can bind to the promoter fragment with W-box motifs, but fail in binding to the promoter fragment with the mutated W-box motifs or without W-box motifs.Therefore, ThWRKY2 and ThWRKY8 are the up-stream regulators of ThERF1, and can regulate the expression of ThERF1 by binding to the W-box motifs in its promoter .