水稻OsPIP1;2植物过表达及亚细胞定位载体的构建

Construction of OsPIP1;2 Over-expression Vector and Transient Vectors of Subcellular Localization

水稻质膜内在蛋白OsPIPs是细胞水分运输的关键蛋白,与多种重要物质的运输交换有关,并在生长发育的各个阶段发挥重要作用。为了挖掘水稻OsPIP1;2基因的功能和应用,本研究利用电子克隆从提取的水稻叶片RNA中成功克隆了OsPIP1;2CDS序列,将其连接到植物表达载体pCAMBIA1301上,构建了35S启动子驱动的OsPIP1;2过表达载体,并成功将其导入到农杆菌EHA105中。此外,为了更全面的了解OsPIP1;2蛋白的亚细胞定位情况,本研究将不同的荧光蛋白分别融合到OsPIP1;2的N端和C端,构建了两个亚细胞定位载体。以上载体的构建,为深入了解和研究OsPIP1;2在水稻中的功能打下了基础。

Rice Plasma membrane intrinsic proteins （OsPIPs） play a key role in water transport and exchange of other substrates across cells, and are involved in numerous physiological processes in plant growth and development. In this study, to explore the function and potential application of OsPIP1;2, CDS of OsPIP1;2 was cloned from rice leaf RNA using insilico cloning and introduced to the Plant expression vector pCAMBIA1301 to construct a 35S driven OPIP1;2 over-expression vector which was successfully transformed into A. tumefaciens EHA105. Meanwhile, two different fluorescent proteins was fused to N and C termini of OsPIP1;2, respectively, to construct two transient expression vectors for comprehensive analysis of OsPIP1;2 subcellular localization. Construction of those vectors lays a foundation for further study and understanding of OsPIP1;2 physiological functions in rice.