中文摘要：

为进一步提高外源基因在水稻悬浮细胞中的表达水平,以花椰菜花叶病毒（Ca MV）35S启动子为对照,克隆了玉米泛素蛋白启动子（Ubi）、水稻肌动蛋白启动子（Actin）以及水稻Oscc1启动子,以GFP为报告基因,通过启动子串联的方式,构建了10个植物表达载体：p CAMBIA 1304 35S-GFP、Ubi-GFP、Actin-GFP、Oscc1-GFP、35S/Ubi-GFP、35S/Actin-GFP、35S/Oscc1-GFP、Ubi/Actin-GFP、Ubi/Oscc1-GFP、Actin/Oscc1-GFP,采用根瘤农杆菌介导的方法,将表达载体导入日本晴胚性愈伤,经悬浮培养分别获得转基因的悬浮细胞系,利用荧光显微镜、酶标仪检测GFP荧光强度,用Western blot检测GFP蛋白质的表达水平。结果发现串联启动子明显强于单个启动子驱动的GFP蛋白质表达水平。在10个表达载体中,35S/Ubi、Ubi/Actin和Ubi/Oscc1是3种优化的启动子组合,其中Ubi/Oscc1串联启动子驱动GFP的表达水平最高,约为对照35S启动子的3倍。

英文摘要：

To improve the level of expression of exogenous gene（green fluorescent protein gene,GFP） in rice suspension cells,ten expression vectors containing GFP reporter gene were constructed as follows using cauliflower mosaic virus promoter（Ca MV） 35S（CK）,the promoter of ubiquitin（Ubi） from maize,and the promoters of actin（Actin） and Oscc1 from rice as follows： p CAMBIA1304 35S-GFP,Ubi-GFP,Actin-GFP,Oscc1-GFP,35 S/Ubi-GFP,35 S/Actin-GFP,35 S/Oscc1-GFP,Ubi/Actin-GFP,Ubi/Oscc1-GFP,and Actin/Oscc1-GFP.Then by applying agrobacterium tumefaciensmediated method,ten expression vectors mentioned above were transformed into Nipponbare rice suspension cells to obtain transgenic Nipponbare rice suspension cells.Fluorescence microscopy and microplate reader were used to detect GFP fluorescence intensity,and GFP expression levels were detected by western blotting.The results showed that tandem promoter driven GFP expression were significantly greater than a single promoter-driven ones.In ten expression vectors,35 S/Ubi,Ubi/Actin and Ubi/Oscc1 were three optimized promoter combinations,among which Ubi/Oscc1 tandem promoter drove the highest expression level of GFP,three times more than the Ca MV35 S promoter.