Regulatory system of gene expression by novel artificial anti-gene chemicals in plants

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[aim] Development of suppression method of gene function by anti-gene chemicals (S-oligo).[experiments]1. Movement of S-oligo in plant cells.2. Suppression of reporter gene function by S-oligo.3. Suppression of transcription factor gene by anti-sense RNA method.[Results and discussion]1. Total DNA was isolated from cultured tobacco cells (BY2) that were cultured in liquid medium containing S-oligo. It was confirmed with Southern blotting analysis and confocal laser scanning microscopy analysis that S-oligo was existence in the BY2 cells.2. We could not detect tie suppression of gene function by S-oligo. Quantitative analysis with in vitro translation system is necessary as next experiment.3. We have isolated a tobacco Ntlim1, as a trans factor binding to a PAL-box motif of the prxC2 promoter. We found that antisense expression of NtUm1 in transgenic plants carrying the prxC2 promoter : GUS chimeric construct decreased not only tie level of tie basal and the wounded-induced expression of tie GUS reporter gene but also the extent of tie wound indudbility of tie prxC2 promoter itself. This result indicates that Ntlim1 is required for the basal level of prxC2 promoter activity as well as its upregulation under wound stress.