HISTOCHEMICAL ANALYSIS OF CAMV 35S PROMOTER-BETA-GLUCURONIDASE GENE-EXPRESSION IN TRANSGENIC RICE PLANTS
HISTOCHEMICAL ANALYSIS OF CAMV 35S PROMOTER-BETA-GLUCURONIDASE GENE-EXPRESSION IN TRANSGENIC RICE PLANTS
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DOI：
10.1007/bf00017828
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发表时间：
1990-10-01
影响因子：
5.1
通讯作者：
HALL, TC
中科院分区：
文献类型：
--
作者：
BATTRAW, MJ;HALL, TC
花椰菜花叶病毒启动子通常用于驱动转基因植物中嵌合基因的转录，包括谷物。为了确定启动子在谷类植物中起作用的组织和细胞类型，对含有CaMV 35S启动子/GUS嵌合基因的转基因水稻植株进行了GUS活性分析。通过未切割和内切DNA的凝胶印迹杂交分析，证实了低拷贝数的35S/GUS嵌合基因插入到由电穿孔原生质体再生的植株的染色体DNA中。定量测量表明，水稻叶片中的GUS活性约为烟草叶片中的10倍[8]，而水稻根中的GUS活性与已报道的烟草根中的GUS活性相似。GUS活性的组织化学定位证实，CaMV 35S启动子在叶表皮、叶肉细胞和维管束细胞中发挥作用。它在根的皮层和维管柱中也很活跃，但在根的表皮中只有少量的活跃。在稳定转化的水稻植株分化组织中GUS活性的分布和水平基本相似，这表明CaMV 35S启动子作为转基因单子叶和双子叶植物基因活性研究的阳性对照是有价值的。
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 [8] 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 differentiatied 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.