CAREER: Aqueous Two-Phase Separation of DNA Using PNA-Conjugated Amphiphiles

事业：使用 PNA 共轭两亲物对 DNA 进行水相两相分离

• 批准号: 0093538
• 负责人: James Schneider
• 金额: $ 37.5万
• 依托单位: Carnegie-Mellon University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0093538&HistoricalAwards=false
• 关键词: CAREER; Aqueous; Two; Phase; Separation; CAREER; Aqueous; Two; Phase; Separation

The PI proposes experiments to identify ATPS polymers and operating conditions so that tagged DNA will strongly partition to one phase, leaving polar proteins and untagged DNA in the other phase. By proper molecular design, the PNA amphiphile will dissociate from the DNA product upon heating the system above the melting transition temperature (Tm) of the PNA-DNA recognition complex. He will also describe a series of theoretical and experimental techniques that will be used to design PNA amphiphiles so that they specifically bind plasmid DNA, but have Tm in a useful range for ATPS. In addition to the development of a useful biochemical process, molecular-level design of the PNA amphiphile-DNA binding complex proposed here should also shed light on the contribution of intermolecular forces to the structure and function of biomolecular materials.His education plan aims to cultivate molecular-level insight in the chemical engineering curriculum by a series of short modules to be assigned in core chemical engineering courses. Molecular modeling and experimental results of the research proposed here will be integrated into the modules, which are designed to predict fundamental engineering properties such as thermal conductivity, viscosity, and vapor-liquid equilibrium from molecular parameters. Another educational effort he proposes is a two-week mini-course for a colloid/interfacial engineering course that emphasizes the connection between biotechnology and colloid science.

PI提出了实验以鉴定ATP聚合物和工作条件，以便标记的DNA将强烈分配到一个相中，在另一阶段留下极性蛋白和未标记的DNA。通过适当的分子设计，PNA两亲物将在加热PNA-DNA识别复合物的熔化过渡温度（TM）上加热系统后与DNA产物分离。他还将描述一系列理论和实验技术，这些技术将用于设计PNA两亲物，以便它们专门结合质粒DNA，但对ATP的TM处于有用的范围内。除了开发有用的生化过程外，这里提出的PNA Amphile-DNA结合复合物的分子水平设计还应阐明分子间力对生物分子材料的结构和功能的贡献。他旨在通过化学工程培养一系列工程的分子工程，旨在培养一系列的工程，以培养一系列的工程，以培养一系列的工具。这里提出的研究的分子建模和实验结果将集成到模块中，这些模块旨在预测分子参数的基本工程特性，例如导热率，粘度和蒸气 - 液体平衡。他提出的另一项教育努力是为期两周的迷你课程，用于胶体/界面工程课程，强调生物技术与胶体科学之间的联系。