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

[aim]抗基因化学品（S-oligo）抑制基因功能方法的开发。[实验]1. S-oligo在植物细胞中的运动. S-oligo对报告基因功能的抑制作用反义RNA法抑制转录因子基因表达。[结果与讨论]1。从在含有S-oligo的液体培养基中培养的培养的烟草细胞（BY2）中分离总DNA。Southern印迹分析和激光共聚焦显微镜分析证实，BY2细胞中存在S-oligo.我们无法检测到S-oligo对基因功能的抑制。体外翻译系统的定量分析是下一步实验的必要条件.我们已经分离出烟草Ntlim 1，作为一个反式因子结合到PAL盒基序的prxC 2启动子。我们发现，在转基因植物携带的prxC 2启动子：GUS嵌合构建体的NtUm 1的反义表达不仅降低了领带的基本水平和领带GUS报告基因的伤口诱导表达，但也领带prxC 2启动子本身的领带伤口诱导的程度。该结果表明，Ntlim 1是prxC 2启动子活性的基础水平以及其在伤口应激下的上调所必需的。

[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.