水稻OsRhoGDI2基因RNA干扰载体的构建

Construction of the RNAi Vector of Rice OsRhoGDI2 Gene

水稻OsRhoGDI2是通过酵母双杂交筛选到的小G蛋白Rho家族成员OsRacD互作蛋白的编码基因,为了研究OsRhoGDI2与OsRacD的相互作用及其在水稻发育中的功能联系,本研究基于序列比对的提示,选择了OsRhoGDI2基因长度为285 bp的特异区段,分别以正向和反向插入中间载体pKANNIBAL中,并亚克隆到受控于CaMV 35S启动子的植物表达载体pART27上,获得了OsRhoGDI2基因的RNA干扰载体,为利用水稻转基因技术鉴定该基因在水稻发育中的功能,以及与OsRacD基因的功能联系奠定基础.

By yeast two-hybrid screening,rice OsRhoGDI2 gene is isolated as the partner of small GTP-binding protein OsRacD which belongs to Rho family.Based on the sequence alignments,the 285bp gene-specific fragment of OsRhoGDI2 is inserted into pKANNIBAL vector in forward and reverse direction respectively,then subcloned into binary plant expression vector pART27,resulting in the RNAi vector pART27-OsRhoGDI2,facilitating further study of the interaction between OsRhoGDI2 and OsRacD,and also its function in rice development.