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）允许在哺乳动物细胞中系统分析蛋白质表达的方法将对阐明这种蛋白质的作用产生重大影响，并最终可能为治疗性发育提供新的靶标。研究人员建议根据蛋白质捕获来设计新工具，以针对哺乳动物细胞中的蛋白质发现和功能分析。蛋白质陷阱系统由具有编码蛋白质标签的人工外显子的移动遗传元件（蛋白质陷阱载体）组成。在插入蛋白质陷阱载体在细胞基因的内含子中后，蛋白质标签序列将在成熟的mRNA中保守。翻译将导致蛋白质标签融合到被困蛋白的氨基和羧基末端部分。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特定经验信息。带有标记蛋白的细胞系也将使自己研究这种蛋白质的空间分布，以响应特定的药物以及细胞分化和发育期间。