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
作者：刘盈盈(四川大学生命科学学院 生物资源与生态环境教育部重点实验室)；刘 宇(四川大学生命科学学院 生物资源与生态环境教育部重点实验室)；黄 奎(四川大学生命科学学院 生物资源与生态环境教育部重点实验室)；姚润东(四川大学生命科学学院 生物资源与生态环境教育部重点实验室)；王健美(四川大学生命科学学院 生物资源与生态环境教育部重点实验室)；李旭峰(四川大学生命科学学院 生物资源与生态环境教育部重点实验室)；杨毅(四川大学生命科学学院 生物资源与生态环境教育部重点实验室)
Author：LIU Ying-Ying(Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University)；LIU Yu(Key Laboratory Bio-resources and Eco-environment of Ministry Education,College of Life Sciences,Sichuan University)；HUANG Kui(Key Laboratory Bio-resources and Eco-environment of Ministry Education,College of Life Sciences,Sichuan University)；YAO Run-Dong(Key Laboratory Bio-resources and Eco-environment of Ministry Education,College of Life Sciences,Sichuan University)；WANG Jian-Mei(Key Laboratory Bio-resources and Eco-environment of Ministry Education,College of Life Sciences,Sichuan University)；LI Xu-Feng(Key Laboratory Bio-resources and Eco-environment of Ministry Education,College of Life Sciences,Sichuan University)；YANG Yi(Key Laboratory Bio-resources and Eco-environment of Ministry Education,College of Life Sciences,Sichuan University)
收稿日期：2018-02-09 年卷（期）页码：2018,55(6):1301-1306
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：钙依赖蛋白激酶12；ABA响应原件结合因子4；氨基酸序列比对；酵母双杂交实验；双分子荧光互补实验
Key words：CPK12； ABF4； Sequence alignment of amino acid； Yeast two-hybrid system； BIFC
基金项目：国家转基因重大专项(2016zx08009003-002)
中文摘要
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. 。
英文摘要
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.
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