DNA-Protein binding assays

DNA-蛋白质结合测定

• 批准号: 186118-2008
• 负责人: Le, Chris
• 金额: $ 5.25万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2011
• 资助国家: 加拿大
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=476552
• 关键词: DNA; Protein; binding; assays

Interactions between DNA and proteins are responsible for many biological processes in the cell. The affinity and specificity of these interactions can be exploited for bioanalytical assays that make use of DNA-protein binding. The primary objective of this research is to develop novel bioanalytical technology combining DNA-protein affinity recognition, capillary electrophoresis separation, and laser induced fluorescence detection. Both established and novel affinity probes, including antibodies, DNA damage recognition proteins, and short synthetic DNA molecules (aptamers), will be examined for their ability to bind with the target molecules. To assay for DNA damage, for example, DNA will be isolated from cells and incubated with a fluorescent probe recognizing the damage. The complex of the affinity probe with the damaged DNA will be separated from other molecules by capillary electrophoresis and its concentration will be quantified by laser induced fluorescence.

DNA和蛋白质之间的相互作用负责细胞中的许多生物过程。这些相互作用的亲和力和特异性可用于利用DNA-蛋白质结合的生物分析测定。本研究的主要目的是发展一种结合DNA-蛋白质亲和识别、毛细管电泳分离和激光诱导荧光检测的新型生物分析技术。已建立的和新的亲和探针，包括抗体，DNA损伤识别蛋白，和短的合成DNA分子（适体），将检查其与靶分子结合的能力。例如，为了检测DNA损伤，将从细胞中分离DNA，并与识别损伤的荧光探针一起孵育。亲和探针与受损DNA的复合物将通过毛细管电泳与其他分子分离，并且其浓度将通过激光诱导荧光定量。