Quantitating In Vitro Selection

定量体外选择

• 批准号: 0403139
• 负责人: Steven Benner
• 金额: $ 24万
• 依托单位: University of Florida
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2005-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0403139&HistoricalAwards=false
• 关键词: Quantitating; Vitro; Selection

Professor Benner will lead a project that combines organic synthesis, biological chemistry, in vitro evolution (lVE) and univariate statistical analysis that will develop the physical organic chemical tools needed to quantitate the impact of adding chemical functionality (amino, thiol, imidazole, peptides, and metal chelating groups) via covalent attachment to catalytic DNA molecules. This work will first address the paradox that arises from the observation that endowing DNA and RNA libraries with additional functionality evidently improves the catalytic potential of nucleic acids by only a factor of two to ten, not by the orders of magnitude expected from standard Structure Theory in Organic Chemistry. A set of five hypotheses that account for the orders-of-magnitude discrepancy between expectation and observation will be tested, including models that hold (a) that our view of the role of functionality in catalysis is naive, (b) that the functional endowment of natural DNA (phosphates and hydrogen bond donating and accepting groups) are sufficient for catalysis in general, (c) that the functionality recruited non-covalently by nucleic acids from solution (in particular, divalent cations such as Mg++) overwhelms the contribution of covalently linked functionality, (d) that lVE experiments lose the best catalysts, and (e) that more than one type of functional group is needed before the expected large benefit from functionality is seen. This will require careful assessment of the kinetic order of the reaction being effected by the selected DNA molecules (first order, unimolecular, with the catalytic step rate determining?).With this Award, the Organic and Macromolecular Chemistry Program (OMC) and the Molecular Biochemistry Program in the Division of Molecular and Cellular Biosciences (MCB) will support the research of Professor Steven A. Benner of the University of Florida. Professor Benner's work is expected to have broad impact. From a practical perspective, we may learn how to truly get "catalysis on demand" from IVE experiments, useful for everything from biomedicine to environmental remediation. From a scientific perspective, we will understand in greater depth the possibilities of single biopolymer systems playing a role in the origin of life. From a methodological perspective, he will develop tools that permit IVE to explore the distribution of chemical properties in "structure space" defined by a DNA sequence. And, from a theoretical perspective, the work may end up altering, perhaps dramatically, our global view of the relationship between functionality and reactivity in nucleic acids.

Benner教授将领导一个项目，该项目结合了有机合成，生物化学，体外进化（LVE）和单变量统计分析，该项目将开发定量化学功能（氨基，硫醇，咪唑，肽，肽和金属螯合组）的影响所需的物理有机化学工具，这是通过共聚型DNA Molecules to dna Molecules covalent covalent conterment。这项工作将首先解决源自观察到的悖论，即赋予DNA和RNA库具有其他功能的悖论显然将核酸的催化潜力提高了两到十倍，而不是由有机化学中标准结构理论预期的数量级。将测试一组五个假设，这些假设解释了期望和观察之间的稳定性差异，包括（a）持有的模型，即我们对功能在催化中的作用的看法是天真的，（b）通过自然DNA（磷酸盐和水力键合成组）的功能性赋予型号，不足以供应型蛋白（C型），而不得以斗争的速度（C），（c）在一般情况下（c），（c），（c）的功能（c），c cy coperication contecity（c），C）在（C）中（C），C）（C），C）（C），C）（C），C）（C），C）解决方案（尤其是Mg ++等二价阳离子）压倒了共价连接的功能的贡献，（d）LVE实验会丢失最佳催化剂，并且（e）在看到预期的大量益处需要多种功能组之前，才能看到功能的大益处。这将需要仔细评估由选定的DNA分子（一阶，单分子的一阶，催化步骤速率决定的一阶）。通过该奖项，有机和大分子化学计划（OMCC）和分子生物化学计划在分子和蜂窝生物学家的分部中支持MC的分子生物化学计划（MC）。 预计本纳教授的工作将产生广泛的影响。从实际的角度来看，我们可以学习如何从IVE实验中真正获得“按需催化”，这对从生物医学到环境修复的所有方面都有用。从科学的角度来看，我们将更深入地了解单个生物聚合物系统在生活起源中发挥作用的可能性。从方法论的角度来看，他将开发工具，使IVE可以探索由DNA序列定义的“结构空间”中化学性质的分布。而且，从理论的角度来看，这项工作可能最终改变了我们对核酸功能与反应性之间关系的全球视野。