拟南芥钙依赖蛋白激酶CPK12与bZIP转录因子ABF4相互作用研究

The research on interaction between Arabidopsis Calcium-dependent kinanse CPK12 and bZIP transcription factor ABF4

为分析拟南芥钙依赖蛋白激酶CPK12在不同物种中序列的相似性,并研究CPK12与ABA信号通路的关系,本研究对不同物种CPK12氨基酸序列进行比较,以及通过体内和体外实验验证CPK12与ABF4之间是否存在直接相互作用关系.通过对拟南芥(Arabidopsis thaliana),油菜(Brassica napus)与荠菜(Capsella rubella)等物种的同源氨基酸序列进行比较,获得了它们的亲缘关系.利用酵母双杂交系统,将CPK12与ABF4共转酵母后,在四缺型培养基上能够正常生长,这说明CPK12与ABF4在体外有相互作用.同时,利用双分子荧光互补实验,CPK12与ABF4共转烟草后能够在激发光下观察到烟草细胞核中出现明显荧光信号,这说明CPK12与ABF4在植物体内也存在相互作用,为CPK12参与ABA信号途径提供了直接证据.

In order to analyze the sequence similarity of the Arabidopsis calcium-dependent protein kinase CPK12 in different species and the function of CPK12 in the ABA signaling, this study carried on the amino acid sequence alignment of CPK12 among different species, and focused on the direct interaction between CPK12 and ABF4 in vivo and in vitro . The amino acid sequence alignment was conducted to analyze CPK12 homology among the Arabidopsis thaliana , Brassica napus, Capsella rubella , etc. In addition, CPK12 and ABF4 were co-transfected through the yeast two-hybrid system. The result showed that yeasts could grow normally on SD （-His/Ade Leu/Trp） agar medium, indicating that CPK12 interacted with ABF4 in vitro . In the bimolecular fluorescence complementary assay, the obvious yellow fluorescence signal could be observed in the nucleus when CPK12 and ABF4 were co-transfected into tobacco. The result also demonstrated that CPK12 interacted with ABF4 in vivo . All these provided direct evidence that CPK12 was involved in the ABA signaling.