拟南芥H^+-ATPase AHA11蛋白与钙结合蛋白CBL10的互作关系分析  

Interaction Analysis of Arabidopsis H^+-ATPase AHA11 and Calcium-binding Protein CBL10

作　　者：杨莹[1] 周扬[1] 洪莎[1] 江行玉[1] 

机构地区：[1]海南大学农学院/海南省热带生物资源可持续利用重点实验室,海口570228

出　　处：《基因组学与应用生物学》2016年第4期957-962,共6页Genomics and Applied Biology

基　　金：国家自然科学基金(31260218;31160185)资助

摘　　要：采用分子克隆技术,从拟南芥中分离到一个质膜H^+-ATPase基因At AHA11和一个钙结合蛋白基因At CBL10。采用酶切连接的方法构建At AHA11-C-ter-p GADT7和At CBL10-p GBKT7酵母双杂交载体,用PEG/Li Ac法将At AHA11-C-ter-p GADT7转化到酵母菌Y187中,At CBL10-p GBKT7转化到酵母菌Y2HGold中。发现融合的二倍体酵母(At AHA11-C-ter-p GADT7×At CBL10-p GBKT7)可以在酵母选择性培养基SD-A-H-L-T+X-α-Gal+Ab A中生长。说明At CBL10蛋白可与AHA11蛋白的C末端互作,便于我们进一步研究它们的调节位点,为深入探讨CBL10调控细胞膜AHA11蛋白的功能提供一个技术平台。An H~＋-ATPase gene At AHA11 and a calcium-binding protein gene At CBL10 were isolated from Arabidopsis by molecular cloning technique. The yeast two hybrid expressing vectors At AHA11-C-ter-p GADT7 and At CBL10-p GBKT7 were constructed using enzyme digestion and ligation methods, and then transformed into the Y187 and Y2 HGold yeast strains by PEG/Li Ac methods, respectively. The diploid yeast fused by the transformed Y187 and Y2 HGold yeast strain could grow on the selective yeast plates（SD-A-H-L-T＋X-α-Gal＋Ab A）. The results illustrated that At CBL10 protein could interact with the C-terminal of AHA11 protein, which was convenient for studying the regulation sites of At CBL10 protein, and found a technical platform for researching the mechanism of CBL10 protein regulated plasma membrane AHA11 protein.

关 键 词：质膜H^+-ATPASE 钙结合蛋白 酵母双杂交 拟南芥 

分 类 号：Q946[生物学—植物学]