GENE REGULATION OF HUMAN CONE TRANSDUCIN ALPHA SUBUNIT

人锥转导蛋白α亚基的基因调控

基本信息

批准号：
3265932
负责人：
SHAO-LING FONG
金额：
$ 22.55万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-03-01 至 1996-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3265932
关键词：
DNA footprinting chemical binding complementary DNA cone cell gel mobility shift assay gene expression genetic regulation human genetic material tag molecular cloning molecular site nucleic acid sequence polymerase chain reaction protein sequence retina retinoblastoma transcription factor transducin transfection

项目摘要

The long-term goal of the proposed study is to understand the control of gene expression in photoreceptor cells. This proposal is designed to use the human cone transducin gene as a model for cone photoreceptor-specific gene expression. The components that are involved in the regulation mechanism are cis-acting elements and trans-acting factors. The cis- acting elements will be defined by transfecting retinoblastoma cells with an expression vector containing a reporter gene and a relevant DNA sequence. Trans-acting factors in the nuclei of retinoblastoma cells and in adult human retinas will be identified by DNase 1 footprinting and mobility shift assays. Protein-binding sites on cis-acting elements will be determined by methylation interference assays. The trans-acting factors identified will be cloned by screening a retina expression library with a specific cis-acting element. The identity of the trans- acting factors will be further confirmed by determining the partial amino acid sequences from the proteins or from the peptides. This work will generate new insights into gene expression in photoreceptors, and will provide a basis for the study of inherited retinal diseases at the molecular level.

拟议的研究的长期目标是了解对感光细胞中的基因表达。该建议旨在使用人锥转素基因作为锥形感受器特异性的模型基因表达。法规中涉及的组件机制是顺式作用元素和跨性因素。顺式 - 作用元件将通过与包含报告基因和相关DNA的表达矢量顺序。视网膜细胞细胞核中的跨作用因子和在成年人类的视网膜中，将通过DNase 1足迹识别移动转移测定法。顺式作用元件上的蛋白质结合位点将通过甲基化干扰测定法确定。跨性别确定的因素将通过筛选视网膜表达来克隆具有特定顺式作用元素的库。转移的身份通过确定部分氨基，将进一步确认作用因素来自蛋白质或肽的酸序列。这项工作将对感光体中的基因表达产生新的见解，并将为研究遗传性视网膜疾病的研究提供了基础分子水平。