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植物类钙调磷酸酶B蛋白CBLs (Calcineurin B-like proteins)和CBL互作蛋白激酶CIPKs (CBL-interacting protein kinases)在应答非生物逆境的钙信号系统中起着重要作用.为了研究小麦TaCIPK3的互作调控网络,以普通六倍体小麦(Triticum aestivum)品种矮抗58的叶片cDNA为模板,PCR扩增获得TaCIPK3的编码序列.TaCIPK3基因开放阅读框(ORF)长l 348 bp,编码447个氨基酸.生物信息学分析显示TaCIPK3的N-端催化结构域含有CIPK家族蛋白的典型激活环,C-端调节域含有该家族高度保守的24个氨基酸NAF基序,具有丝氨酸-苏氨酸蛋白激酶的结构特征.利用酵母双杂交体系对TaCIPK3与TaCBL1、TaCBL2、TaCBL3、TaCBL4、TaCBL6、TaCBL7、TaCBL9进行相互作用鉴定,发现分别转化有pGADT7-TaCIPK3×pGBKT7-TaCBL 1、pGADT7-TaCIPK3×pGBKT7-Ta-CBL2、pGADT7-TaCIPK3×pGBKT7-TaCBL3和pGADT7-TaCIPK3 ×pGBKT7-TaCBL4的二倍体酵母菌能够在二缺培养基SD/-Trp/-Leu平板上长出白色菌落,同时这些二倍体菌株在四缺培养基SD/-Leu/-Trp/-Ade/-His/X-α-Gal/AbA上能够长出淡蓝色菌落,表明下游的4个报告基因ADE2、HIS3、MEL1和AUR1-C被激活,因此说明TaCIPK3与TaCBL1、TaCBL2、TaCBL3和TaCBL4之间存在互作.本研究结果对进一步研究TaCIPK3的生物学功能以及与TaCBLs的互作网络具有一定的参考作用.“,”Calcineurin B-like proteins (CBLs) and CBL-interacting protein kinases (CIPKs) play important roles in the Calcium signaling system that is in response to abiotic-stresses.In order to study the interaction network of TaCIPK3 in wheat,the coding sequence of TaCIPK3 gene was amplified from wheat cultivar AiKang58,using leaf cDNA libraries as PCR template.The open reading frame (ORF) of TaCIPK3 gene was 1 348 bp in length,which encoded 447 amino acids.Bioinformatics analysis revealed that the TaCIPK3 protein contained a typical activation loop of CIPK proteins at its N-terminal kinase catalytic domain,a NAF motif consisted of 24 highly conserved amino acids at the C-terminal regulatory domain,which had the structural characterization of serine threonine protein kinase.Yeast Two-Hybrid assays were used to test the interactions between TaCIPK3 and TaCBL1,TaCBL2,TaCBL3,TaCBL4,TaCBL6,TaCBL7,TaCBL9.Yeast strains harboring pGADT7-TaCIPK3 ×pGBKT7-TaCBL1,pGADT7-TaCIPK3×pGBKT7-TaCBL2,pGAD-T7-TaCIPK3×pGBKT7-TaCBL3 and pGADT7-TaCIPK3 ×pGB-KT7-TaCBL4,respectively,could cultivate white plaques on SD/-Trp/-Leu plates.They also showed blue plaques on SD/-Leu/-Trp/-Ade/-His/X-α-Gal/AbA plates,indicating that the four downstream reporter genes A DE2,HIS3,MEL1 and A UR1-C were activated.Therefore,it was concluded that TaCIPK3 could interact with TaCBL1,TaCBL2,TaCBL3 and TaC BL4.These results might facilitate our understanding of the function of TaCIPK3 and its networking with TaCBLs.