Factors Involed in Chloroplast RNA Editing

描述（由申请人提供） 这个项目的长期目标是定义RNA：RNA和RNA：蛋白质 参与RNA编辑的相互作用。在人类中，编辑对RNA至关重要 处理几种疾病相关基因的表达，包括 威尔姆s肿瘤易感基因，谷氨酸受体亚单位B，和 载脂蛋白B。这项研究将阐明直接胞苷 尿苷编辑和增加我们对细胞器编辑的理解 机制等 在研究期间，转基因和生物化学技术将 用于鉴定与叶绿体rpoB-I编辑相互作用的分子 绝佳的价钱为了研究RNA二级结构在编辑中的作用，rpoB-I将在 与反义序列融合并在转基因烟草叶绿体中表达。 将进行电泳迁移率变化测定以鉴定叶绿体 对体外转录的rpoB-I编辑具有结合特异性的蛋白质 网站. RNA：蛋白质相互作用的性质将使用 修改编辑序列。推测的蛋白质的纯化和表征 编辑蛋白质将通过亲和方法来促进。的功能 筛选将用于分离编码编辑因子的基因。

DESCRIPTION (provided by applicant) The long-term objective of this project is to define RNA:RNA and RNA:protein interactions involved in RNA editing. In humans, editing is critical for RNA processing in the expression of several disease-related genes including the Wilm?s tumor susceptibility gene, glutamate receptor subunit B, and apolipoprotein B. This research will elucidate the factors that direct cytidine to uridine editing and increase our understanding of organellar editing mechanisms. During the fellowship period, transgenic and biochemical techniques will be used to identify molecules that interact with the chloroplast rpoB-I editing site. To study the role of RNA secondary structure in editing, rpoB-I will be fused to antisense sequences and expressed in transgenic tobacco chloroplasts. Electrophoretic mobility-shift assays will be performed to identify chloroplast proteins with binding specificity for in vitro transcribed rpoB-I editing sites. The nature of the RNA:protein interaction will be examined using modified editing sequences. The purification and characterization of putative editing proteins will be facilitated by an affinity approach. A functional screen will be utilized for the isolation of genes encoding editing factors.