Multiwell colorimetric assay for DNA repair protein.
DNA 修复蛋白的多孔比色测定。
基本信息
- 批准号:6582810
- 负责人:
- 金额:$ 9.94万
- 依托单位:
- 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:2002-09-30 至 2003-09-30
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
DESCRIPTION (provided by applicant): DNA damages are key events during carcinogenesis. Deficiencies in repairing damaged DNA represent a poor prognosis during tumorigenesis and/or cancer treatment. Besides monitoring DNA damage formation, it would be helpful to monitor DNA repair protein activity. We propose to develop a new technology that allows for high-throughput and fast analysis of the most common DNA repair proteins. We will use a damaged DNA probe immobilized onto a 96-well microplate to capture DNA repair proteins in an extract from cells or tissues. Six biotinylated oligonucleotides containing different DNA damage types will be synthesized. The DNA-protein binding conditions will be developed by using purified recombinant DNA repair proteins. Antibodies raised specifically against DNA repair proteins will identify the bound protein and yield a quantitative result in two hours. Finally, the procedure will be applied to relevant samples such as extracts from damaging agent-treated cell lines and disease models. Such assay will help elucidate the molecular mechanisms of the DNA damage and repair pathways. It would also provide scientists and clinicians with a unique tool to answer seminal questions in risk assessment in a fraction of the time normally required.
描述(由申请人提供):DNA损伤是致癌过程中的关键事件。修复受损DNA的缺陷代表肿瘤发生和/或癌症治疗期间的不良预后。除了监测DNA损伤形成外,监测DNA修复蛋白的活性也将是有帮助的。我们建议开发一种新技术,可以高通量和快速分析最常见的DNA修复蛋白。我们将使用固定在96孔微孔板上的受损DNA探针来捕获细胞或组织提取物中的DNA修复蛋白。将合成六种含有不同DNA损伤类型的生物素化寡核苷酸。将通过使用纯化的重组DNA修复蛋白来开发DNA-蛋白结合条件。针对DNA修复蛋白的特异性抗体将识别结合的蛋白质,并在两小时内产生定量结果。最后,该程序将应用于相关样品,如损伤剂处理的细胞系和疾病模型的提取物。这种检测将有助于阐明DNA损伤和修复途径的分子机制。它还将为科学家和临床医生提供一种独特的工具,以在通常所需时间的一小部分内回答风险评估中的开创性问题。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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{{ truncateString('JACQUES T WEISSMAN', 18)}}的其他基金
High throughput assay for small G protein activation
小 G 蛋白激活的高通量测定
- 批准号:
6932935 - 财政年份:2005
- 资助金额:
$ 9.94万 - 项目类别:
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