农杆菌介导的水稻Waxy基因过量表达初步研究

摘要

采用RT-PCR技术克隆水稻Waxy基因,并构建该基因的过量表达载体,在农杆菌介导下利用建立的水稻遗传转化体系将目的基因转入水稻.转基因水稻经PCR检测及实时定量PCR拷贝数检测均表明外源基因已成功导入水稻,且多数为单拷贝插入.半定量PCR检测结果显示,导入水稻的外源Waxy基因已成功表达,并使得该基因的表达量较对照有明显提高.水稻Waxy基因过量表达载体的成功转化,将为该基因功能的深入研究及水稻直链淀粉合成的调控奠定基础.%To obtain high amylose transgenic rice, the functions of Waxy genes in amylose synthesis in rice were studied. The Waxy gene was cloned by the technique of RT-PCR, the vector of over-expression of Waxy genes was constructed. The target gene was transformed into rice using rice genetic transformation system mediated by Agrobacterium. It showed that exogenous genes had been successfully transformed into rice by the mean of the single-copy insertion by PCR detection and real-time quantitative PCR. The results showed the exogenous Waxy gene had been successfully expressed, and the expression amount was improved obviously than the control. It would become the foundation of the further study of the gene function and regulation of amylose synthesis in rice that the vector of over-expression of Waxy gene was transformed successfully.