| SPS (sucrose phosphate synthase) plays an important role in the sucrose formation during CO2 photo assimilation in the plants. At present, the transcription mechanism of SPS in sugarcane has not yet clear, however, the core region of promote plays a key role in the regulation of eukaryotic gene transcription. It is a fact that the research on the promoter of SPS of sugarcane is particularly important.The functional analysis of the promoter of SPSⅢwas studied from Saccharum Officinarum as the main subject in the research. To explore the mechanism which regulates the tissue-specific of SPS, the expression of reporter gene of GUS (β-glucuronidase) was examined which was directed by different lengths of 5'flanking sequence. Until now, the results are as follows:1. We have already constructed five plasmid vectors of deletion expression.Then we carried out a deletion analysis of the sugarcane SPSⅢpromoter using transgenic tobacco plants by Agrobacterium-mediated leaf disc transformation method. PCR results showed that we got transgenic tobaccos. The results of GUS staining are as follows:(1) It was observed that the gene GUS directed by the fragment downstream to position-1439 expressed in the leaf, stem, root neck, receptacle, sepal, petal, anther, style and in the leaf of seedlings. The results confirmed the promoter activity of 5'sequence of SPSⅢ.(2) No activity was detected of the fragment downstream to position-932.(3) It was observed that the gene GUS directed by the fragment downstream to position-725 expressed only in the base of receptacle and anther.(4) It was also observed that the gene GUS directed by the fragment downstream to position-476 expressed only in the receptacle and the base of sepal.(5) No activity was detected of the fragment downstream to position-216.2. Based on the results above, the 5'-flanking sequences of SPSⅢwere fused to the GUS gene in different lengths to construct three plasmid vector of deletion expression. |
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