Battraw M J, Hall T C
Department of Biology, Texas A&M University, College Station 77843-3258.
Plant Mol Biol. 1990 Oct;15(4):527-38. doi: 10.1007/BF00017828.
The cauliflower mosaic virus promoter is commonly used to drive transcription of chimeric genes in transgenic plants, including the cereals. To determine the tissue and cell types of cereal plants that the promoter functions in, transgenic rice plants containing a CaMV 35S promoter/GUS chimeric gene were analyzed for GUS activity. Insertion of a 35S/GUS chimeric gene at low copy number into chromosomal DNA of plants regenerated from electroporated protoplasts was confirmed by gel blot hybridization analysis of uncut and endonuclease-digested DNA. Quantitative measurement showed that GUS activity was some tenfold higher in rice leaves than in tobacco leaves whereas activities obtained for rice roots were similar to those reported for tobacco roots. Histochemical localization of GUS activity confirmed that the CaMV 35S promoter functions in cells of the leaf epidermis, mesophyll and vascular bundle. It is also active in the cortex and vascular cylinder of the root, but only marginally active in the root epidermis. The generally similar distribution and levels of GUS activity obtained in differentiated tissue of stably transformed rice plants indicates the value of the CaMV 35S promoter as a positive control for studies in gene activity in transgenic monocots and dicots.
花椰菜花叶病毒启动子常用于驱动转基因植物(包括谷类作物)中嵌合基因的转录。为了确定该启动子在谷类植物中发挥功能的组织和细胞类型,对含有花椰菜花叶病毒35S启动子/β-葡萄糖醛酸酶(GUS)嵌合基因的转基因水稻植株进行了GUS活性分析。通过对未切割和经核酸内切酶消化的DNA进行凝胶印迹杂交分析,证实了低拷贝数的35S/GUS嵌合基因插入到了由电穿孔原生质体再生的植物染色体DNA中。定量测量表明,水稻叶片中的GUS活性比烟草叶片高约十倍,而水稻根中的活性与报道的烟草根中的活性相似。GUS活性的组织化学定位证实,花椰菜花叶病毒35S启动子在叶片表皮、叶肉和维管束细胞中发挥功能。它在根的皮层和维管束中也有活性,但在根表皮中活性微弱。在稳定转化的水稻植株的分化组织中获得的GUS活性分布和水平总体相似,这表明花椰菜花叶病毒35S启动子作为转基因单子叶植物和双子叶植物基因活性研究的阳性对照具有重要价值。