Protein trapping tools for mammalian cells

用于哺乳动物细胞的蛋白质捕获工具

基本信息

批准号：
6657391
负责人：
JAKOB REISER
金额：
$ 21.3万
依托单位：
LSU HEALTH SCIENCES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-09-10 至 2004-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant) Methods allowing the systematic profiling of protein expression in mammalian cells will have a major impact in elucidating the role(s) of such proteins and may ultimately provide new targets for therapeutic development. The investigators propose to design novel tools for the discovery and functional analysis of proteins in mammalian cells based on protein trapping. The protein trap system consists of a mobile genetic element (protein trap vector) harboring an artificial exon encoding a protein tag. Upon insertion of the protein trap vector within an intron of a cellular gene, protein tag sequences will be conserved in the mature mRNA. Translation will result in a fusion of the protein tag to both the amino- and carboxyl-terminal parts of the trapped protein. Ideally, the tagged protein will retain the localization properties and functions(s)of the wild-type protein. This approach will facilitate gene expression analyses at the protein level, and can be used to gain specific empirical information about the potential biological functions of such proteins. Cell lines harboring tagged proteins will also lend themselves to study the temporal an spatial distribution of such proteins in response to specific drugs and during cellular differentiation and development.

描述（由申请人提供）允许对哺乳动物细胞中蛋白质表达进行系统分析的方法将对阐明此类蛋白质的作用产生重大影响，并可能最终为治疗开发提供新的靶点。研究人员建议设计新的工具，用于基于蛋白质捕获的哺乳动物细胞中蛋白质的发现和功能分析。蛋白质捕获系统由移动遗传元件（蛋白质捕获载体）组成，其中包含编码蛋白质标签的人工外显子。将蛋白质捕获载体插入细胞基因的内含子后，蛋白质标签序列将在成熟 mRNA 中保守。翻译将导致蛋白质标签与捕获蛋白质的氨基端和羧基端部分融合。理想情况下，标记的蛋白质将保留野生型蛋白质的定位特性和功能。这种方法将促进蛋白质水平的基因表达分析，并可用于获得有关此类蛋白质潜在生物学功能的具体经验信息。含有标记蛋白质的细胞系也有助于研究这些蛋白质响应特定药物以及在细胞分化和发育过程中的时间和空间分布。