两种水稻GDP解离抑制蛋白基因的分离及特征分析

以Rho家族成员OsRacD为诱饵 ,采用酵母双杂交体系 ,分离到两种与OsRacD互作的水稻RhoGDP解离抑制蛋白的基因 ,分别命名为OsRhoGDI1和OsRhoGDI2 .酵母体内结合和GSTpulldown分析结果显示 ,OsRhoGDI1和OsRhoGDI2与野生型和组成型激活的OsRacD都能结合 ,且不依赖GDI的N端部分序列 ;GDI和Rho的结合具有一定的特异性 .两种GDI在水稻根、地上组织和幼穗等多种组织和器官都有表达 ,但在表达特征上存在明显差异 .研究证实 ,在水稻中存在着调控RhoGTPases的GDP解离抑制蛋白基因家族 ,为Rho蛋白功能及相关信号通路的研究奠定了基础

Isolation and Characterization of Two GDP Dissociation Inhibitor Genes from Oryza sativa L

Using Rho family member OsRacD as bait, two rice GDP dissociation inhibitors genes, designated OsRhoGDI1 and OsRhoGDI2 were isolated by yeast two-hybrid. Filter lift assay and GST pull down assay showed that OsRhoGDI1 and OsRhoGDI2 bound both wild type and constitutive activated OsRacD ,which were independent on the N-terminal part sequence. The binding specificity between OsRhoGDIs and Rho was also confirmed. The two OsRhoGDIs were expressed in shoots, roots and panicle with obviously different pattern. The results confirmed that there was a GDP dissociation inhibitors gene family regulating Rho GTPases states in rice, facilitated further study of Rho pathway.