Analysis of mRNA splicing in plant hormone signaling

描述（由申请人提供）： 本研究以拟南芥为模型植物，采用生物信息学驱动的分子遗传学方法，研究选择性剪接作为植物激素信号转导的基因调控机制的作用。高密度寡核苷酸微阵列将用于检查外源生长素、油菜素和赤霉素处理对（i）编码推定的RNA结合蛋白剪接因子的拟南芥基因的表达;和（2）前体mRNA剪接的影响。将鉴定T-DNA突变体插入系中编码受激素调节的推定RNA结合蛋白的基因，并测定激素相关表型变化。将使用RT-PCR、变性高效液相色谱（DHPLC）和测序最终确认进一步分析使用Affytek阵列检测到的推定的剪接调控的可变剪接变体。将选择经证实表现出剪接调控的可变剪接的基因用于开发可变剪接报告基因系统，该系统将在转基因植物中进行实验测试。

DESCRIPTION (provided by applicant): A bioinformatics-driven molecular genetics approach will be used to study the role of alternative splicing as a gene regulation mechanism in plant hormone signaling using the model plant Arabidopsis thaliana. High-density oligonucleotide microarrays will be used to examine the effects of exogenous auxin, brassinolide, and gibberellic acid treatments on (i) the expression of Arabidopsis genes that encode putative RNA-binding protein splicing factors; and (2) pre-mRNA splicing. T-DNA mutant insertion lines will be identified for the genes encoding hormone-regulated putative RNA-binding proteins and assayed for hormone associated phenotypic changes. Putative hormone-regulated alternative splice variants detected using the Affymetrix arrays will be further analyzed using RT-PCR, denaturing high performance liquid chromatography (DHPLC) and final confirmation by sequencing. Genes confirmed to exhibit hormone-regulated alternative splicing will be selected for use in developing an alternative splicing reporter gene system that will be tested experimentally in transgenic plants.