马铃薯ERF转录因子相互作用蛋白质的筛选与鉴定

乙烯响应因子(ERF)是植物所特有的一类转录因子，在植物响应生物及非生物胁迫中起着重要的调节作用。目前，在马铃薯中关于ERF转录因子及与其相互作用的蛋白质的调控机制还不清楚。在前期研究中，我们从马铃薯中克隆了ERF家族的5个基因StERF1-4和Pti4，并通过酵母单杂交系统验证表明马铃薯StERFs能通过结合GCC-box来参与对干旱、NaCl、乙烯、水杨酸和茉莉酸甲酯等非生物胁迫的响应。本研究将在此基础上，用干旱胁迫的马铃薯材料构建酵母杂交文库，然后用酵母单杂交和双杂交技术进一步筛选新的马铃薯ERFs转录因子及与其相互作用的蛋白质，用体外pull-down试验和体内免疫共沉淀(CoIP)方法对筛选得到的相互作用蛋白质进行验证，解析ERFs和其相互作用蛋白共同调控马铃薯抗旱性的机理，阐明转录因子ERF调控马铃薯抗旱性的分子机制，为马铃薯抗旱性的遗传改良提供分子基础。

Ethylene response factor (ERF) is a kind of plant-specific transcription factors which play an important regulatory role in plant response to biotic and abiotic stress. At present, the regulation mechanism of transcription factor ERF and proteins interacting with them is not elucidated. In our previous study, the five genes of ERF family including StERF1-4 and Pti4 were cloned from potato, and the five StERFs can bind with GCC-box to involve in potato response to abotic stress including drought, NaCl, ethylene, sulphosalicylic acid and methyl jasmonate using yeast one-hybrid system. In the present research, yeast hybrid library will be constructed using potato materials under drought stress, and further screened novel potato ERF transcription factors and proteins interacting with them using yeast one-hybrid and yeast two-hybrid systems. The proteins interacting with ERFs will be verified by in vitro pull-down assay and in vivo co-Immunoprecipitation (CoIP). The aim of the research is to analyze mechanism of ERFs and their interacting proteins regulating potato drought resistance, and elucidate molecular mechanism of ERF regulating drought resistance of potato, and provide the molecular foundation for genetic improvement of stress resistance of potato.