利用酵母双杂交系统筛选与黄瓜花叶病毒外壳蛋白互作的寄主蛋白

利用酵母双杂交系统,以黄瓜花叶病毒(Cucumber mosaic virus,CMV)的外壳蛋白(coat protein,CP)为诱饵,从番茄叶片c DNA文库中筛选与其互作的蛋白。结果显示,诱饵载体pBT3-SUC-CMV-CP均能在酵母细胞中正确表达,无自激活活性而且对酵母无毒性;通过对酵母双杂交文库的筛选和回转验证,共获得了98个阳性克隆,分别编码67个可能与CMV-CP相互作用的蛋白,分别参与植物防御反应、光合作用、物质转运、信号转导、能量代谢、氨基酸代谢、细胞壁的形态建成、植物的激素代谢等。本研究结果表明,CMV CP可同时调控寄主的多个代谢过程,在CMV的致病过程中有多重功能。

Host Proteins that Interact with Coat Protein of Cucumber mosaic virus Screened by Yeast Two Hybrid System

To identify host proteins that interact with coat protein(CP)of Cucumber mosaic virus(CMV),yeast twohybrid assay was performed to screen a tomato leaf cDNA library using the CP of CMV as bait.Bait plasmid(pBT3-SUC-CMV-CP)expressing CMV-CP showed no sign of self-activation and was non cytotoxic to the cells.After screening and retesting the interaction in yeast,98 candidate clones encoding collectively 67 proteins from the tomato library displaying specific interaction with CMV CP were identified.These candidate interacting proteins were predicted to function in defense reaction,photosynthesis,material transport,signal transduction,energy metabolism,amino acid metabolism,cell wall morphogenesis,and plant hormone metabolism,which suggest that CMV CP might be a multi-functional protein that perturbs various metabolism pathways of its host during pathogenesis.