Analysis of the Ribonucleases of Arabidopsis thaliana

本研究的主要目的是研究模式植物的RNA降解机制是如何被控制的。具体来说，拟南芥核糖核酸酶（RNase）的区隔化，以及对RNase活性的时空限制将被研究。我们的初步实验使用底物凝胶法鉴定了拟南芥中15种RNase活性。我们计划通过梯度分离和细胞器纯化来研究这些酶的区隔化。与液泡、细胞核、叶绿体和多体相关的rna将被识别。对于一个或多个选定的RNases，将进行蛋白质纯化，并培养抗血清用于进一步分析。初步研究表明，一些rna酶在拟南芥中积累存在差异。因此，在不同的发育阶段和不同的环境条件下，不同器官的RNase谱将被比较。除了对基础知识的贡献外，本研究还将进一步提高拟南芥作为研究单个rna酶功能的遗传模型的潜力。//

The primary aim of this research is to investigate how the RNA degrading machinery of a model plant is controlled. Specifically, the compartmentalization of ribonucleases (RNase in Arabidopsis, and the temporal and spacial constrains on RNase activity will be studied. Our initial experiments have identified 15 RNase activities from Arabidopsis using a substrate-based gel assay. We plan to investigate the compartmentalization of these enzymes by gradient fractionation and organellar purification. RNases associated with vacuoles, nuclei, chloroplasts, and polysomes will be identified. For one or more selected RNases, protein purification will be performed and antiserum will be raised and used for further analyses. Preliminary studies indicate that some RNases accumulate differentially in Arabidopsis. THerefore, the RNase profiles in different organs will be compared at different stages of development, and under different environmental conditions. Beyond it contribution to basic knowledge, this research should enhance the potential use of Arabidopsis as a genetic model to study the functions of individual RNases in the future.//