草莓镶脉病毒(SVBV)启动子的鉴定.pdfVIP

benthamiana to identify promoter activity by conducting the quantitive analysis of promoter activity via gus fluorophotometry method. 3. Histochemical assay of gus activity and observation of gfp fluorescence activity in transient expression Promoter activity was determined by histochemical stain. The result revealed that blue spots were observed in fibro-vascular tissue and partial cortical cell of the stem of N. tabacum plants in which pINT P1, pINT P2, pINT P3, pINT P4, pINT P5 and pINT121 expressed. It demonstrated that all the promoters can drive gene gus to express in the plant cells. The expressing level of gene gus driven by the promoter of pINT P1 was higher than that of 35S promoter of pINT121 according to the gus dyeing intensity strength of the slices. Promoter activity was also analyzed by Fluorescence microscope using gene gfp. The result manifested that green fluorescence coult be observed in fibro-vascular tissue of the stem of N. tabacum plants in which pCHF P1, pCHF P5 and pCHF3 expressed. It indicated that all the three promoters could drive gene gfp to express in the plant cells. The expressing level of gene gfp driven by the promoter of pCHF P1 was higher than that of pCHF3, and the expressing level of gene gfp driven by the promoter of pCHF P1 was higher than that of pCHF3 according to the gfp fluorescence intensity strength of the slices. The gfp fluorescence almost could not be observed in stem slices using non-transgenic tobacco plants injected with A. tumefaciens as negative control. 4. Analysis of gus activity by fluorometric assay in transient expression The result of fluorometric assay showed that pINT P1 shared the highest average relative gus activity (280%) with that of pINT121. The gus activity of pINT P2, pINT P3