用酵母双杂交系统筛选与拟南芥成膜素相关蛋白(PhrIP1)相互作用的蛋白质

Identification of Protein Interaction with Phragmoplastin-interacting Protein1 (PhrIP1) by Yeast Two-hybrird System

用酵母双杂交系统,以pEG202-PhrIP1为诱饵筛选拟南芥AD-cDNA文库,寻找与成膜素相关蛋白(PhrIP1)相互作用的蛋白质。结果表明,在5×106个转化子中得到6个阳性克隆,经测序及体外蛋白质相互作用证实,得到与PhrIP1相互作用的2个靶蛋白,即Ran2小GTP结合蛋白和CIPK6钙调节激酶。

To identify proteins that interact with PhrIP1,the yeast two-hybrid system was taken to screen AD-cDNA library of Arabidopsis thaliana by using PhrIP1(pEG202-PhrIP1) as bait. The results showed that 6 genes were isolated from the library from a yield of 5×106 transformants and two of them encoding proteins could be interacted with PhrIP1 in a protein-protein binding assay in vitro,and Ran2 small GTP binding protein and CBL-interacting protein kinase 6 (CIPK6) by sequencing and blast are determined as the target proteins to study.