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GhZFP1蛋白是从盐胁迫棉花幼苗cDNA文库中分离的一种CCCH型锌指蛋白.初步的生物学功能研究表明,过量表达该基因的转基因烟草耐盐性和抗病性显著提高.为深入研究GhZFP1蛋白的作用机制,构建pGBKT7-m1诱饵表达载体,利用酵母双杂交系统从盐胁迫诱导棉花cDNA文库中筛选与其相互作用的蛋白.通过阳性克隆的表型确定、PCR和限制性内切酶检测以及测序和生物信息学分析,获得9个与诱饵蛋白相互作用的靶蛋白.双分子荧光互补实验证明,GhZFP1与GZIRD19A确实存在互作关系.通过分析这些靶蛋白的已知功能,为研究GhZFP1锌指蛋白的未知生物学功能提供重要信息.
The GhZFP1 protein is a CCCH-type zinc finger protein isolated from the salt-stressed cotton seedling cDNA library.A preliminary biological study showed that salt tolerance and disease resistance of transgenic tobacco over-expressing GhZFP1 were significantly increased.In order to further study the effects of GhZFP1 Protein was constructed and the bait expression vector pGBKT7-m1 was constructed.The yeast two-hybrid system was used to screen the cotton cDNA library for its interaction with salt stress.Through the phenotypic identification of positive clones, PCR and restriction endonuclease assays and Sequencing and bioinformatics analysis to obtain nine target proteins interacting with bait protein.Double-molecule fluorescence complementation experiments confirmed that GhZFP1 did interact with GZIRD19A.Through analyzing the known function of these target proteins, The unknown biological function of the protein provides important information.