Single Molecule Studies of DNA-Cyanine Dye Interactions

DNA-花青染料相互作用的单分子研究

• 批准号: 6317033
• 负责人: JEFFREY T PETTY
• 金额: $ 12.23万
• 依托单位: FURMAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6317033
• 关键词: DNA; atomic force microscopy; cations; chemical bond; chemical chain length; chemical kinetics; chemical stability; chemical structure function; computer simulation; cyanine; fluorescence microscopy; fluorescent dye /probe; hydropathy; intermolecular interaction; model design /development; molecular dynamics; molecular shape; nucleic acid sequence; nucleic acid structure; physical model; solutions; stoichiometry; structural biology; thermodynamics; viscosity

DESCRIPTION (provided by applicant): The cyanine dyes are a class of bicyclic aromatic compounds that are highly fluorescent when associated with DNA. The resulting sensitivity of DNA detection has led to applications ranging from medical diagnosis to molecular biology. The cyanine dyes will also serve as useful models to understand critical features of ligand-DNA interactions because they behave as nonclassical intercalators. This research will use single molecule analysis, as well as macroscopic experimental techniques, to investigate important features of the DNA-cyanine dye interaction from thermodynamic, structural, and kinetic standpoints. I. Equilibrium constant and viscosity measurements will be used to characterize the factors that influence the structure and stability of the DNA-dye complex. Dr. Petty will investigate the role of the structure of the dye, the sequence of the DNA, and the solution environment on the dye-DNA association. II. Dimeric cyanine dyes exhibit strong cooperative interactions when bound to DNA fragments. Atomic force microscopy (AFM) will be used to investigate the structures of these cooperative dye-DNA complexes. The unique capabilities of AFM will allow studies such as the sequence dependence of the cooperativity. III. Because of their favorable fluorescence properties, individual cyanine dye molecules will be studied using single molecule fluorescence microscopy. By eliminating the averaging inherent in bulk studies, new perspectives on the dynamics of DNA-ligand interactions are possible. For example, by measuring the rates of association and dissociation of individual molecules, the role of salt in the mechanism of DNA-ligand interaction will be elucidated. Because of the significance of small molecules and enzymes in DNA regulation, the long-term goal of this project is to use single molecule analysis to provide unique perspectives on these DNA-ligand interactions.

描述（由申请人提供）：氰染料是一类Bicyclic芳香化合物，与DNA相关时，它们高度荧光。这 导致DNA检测的敏感性导致应用范围从 分子生物学的医学诊断。氰胺染料也将作为 了解配体-DNA相互作用的关键特征的有用模型 因为它们的行为是非经典的介入者。这项研究将使用 单分子分析以及宏观实验技术 研究从 热力学，结构和动力学的角度。 I.平衡常数和粘度测量将用于表征 影响DNA-DYE复合物的结构和稳定性的因素。 Petty博士将研究染料结构的作用，即序列 DNA和染料-DNA关联的溶液环境。 ii。二聚体氰染料与 DNA片段。原子力显微镜（AFM）将用于研究 这些合作染料-DNA复合物的结构。独特的功能 AFM将允许研究诸如协作性的序列依赖性。 iii。由于其有利的荧光特性，单个氰染料 将使用单分子荧光显微镜研究分子。经过 消除了批量研究中固有的平均值，对 DNA - 配体相互作用的动力学是可能的。例如，通过测量 单个分子的关联率和解离，盐的作用 在DNA - 配体相互作用的机理中，将阐明。 由于小分子和酶在DNA调节中的重要性，所以 该项目的长期目标是将单分子分析用于 提供有关这些DNA - 配体相互作用的独特观点。