拟南芥蛋白质激酶CARK7与ABA受体RCAR12相互作用的研究

脱落酸(ABA)受体RCARs在ABA信号传导过程中起关键作用,研究ABA受体的相互作用蛋白质对进一步了解植物抗逆的分子机制有重要的意义.本研究通过酵母双杂交发现CARK7与ABA受体RCAR12共转化酵母能在营养缺陷型平板SD/-Trp-Leu-Ade上生长.将CARK7与RCAR12分别克隆到原核表达载体pET-28a(+)和pGEX-6p-1中,构建融合蛋白表达载体,并进行原核表达和亲和纯化.纯化所得蛋白进行GST-pull down实验,结果表明RCAR12能够把CARK7蛋白拉下来.上述结果说明CARK7与RCAR12在体外存在相互作用.进一步通过双分子荧光互补实验发现CARK7与RCAR12在烟草叶片表皮细胞中能产生荧光信号,说明两者在体内同样具有明显的相互作用.

The research on interaction between the Protein kinase CARK7 and the ABA receptor RCAR12 in Arabidopsis

The ABA receptor RCARs play a key role in ABA Signal transduction process, and research on interaction proteins of RCARs has great significance in further understanding the molecular mechanism of plants in adverse environment. The yeast two-hybrid assay and GST-pull down were used to study the interaction between CARK7 and RCAR12. The results showed that there is a significant interaction between CARK7 and RCAR12 in vitro. Then, the bimolecular fluorescence complementation (BiFC) was used to confirm the interaction between CARK7 and RCAR12 in vivo. The BiFC assay indicated that CARK7 was associated with RCAR12 in tobacco epidermal cells. Taken together, these data suggested that CARK7 and RCAR12 closely interact with each other, and illuminating the regulation of the ABA signaling transduction.