Factors Involed in Chloroplast RNA Editing

叶绿体 RNA 编辑涉及的因素

• 批准号: 6700712
• 负责人: CARLA E HEGEMAN
• 金额: $ 4.64万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6700712
• 关键词: DNA directed RNA polymerase; RNA binding protein; SDS polyacrylamide gel electrophoresis; antisense nucleic acid; autoradiography; cell transformation; chloroplasts; computer data analysis; cytidine; denaturing gradient gel electrophoresis; gel mobility shift assay; gene expression; genetically modified plants; intermolecular interaction; ion exchange chromatography; nucleic acid hybridization; nucleic acid structure; plant genetics; polymerase chain reaction; posttranscriptional RNA processing; regulatory gene; tobacco; uridine; yeast two hybrid system

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.

描述（由申请人提供）