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

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 提议进行实验来识别 ATPS 聚合物和操作条件，以便标记的 DNA 强烈分配到一个相，而将极性蛋白和未标记的 DNA 留在另一相。通过适当的分子设计，当将系统加热到 PNA-DNA 识别复合物的熔解转变温度 (Tm) 以上时，PNA 两亲物将从 DNA 产物中解离。他还将描述一系列理论和实验技术，用于设计 PNA 两亲物，使其特异性结合质粒 DNA，但 Tm 处于对 ATPS 有用的范围内。除了开发有用的生化过程之外，这里提出的 PNA 两亲物-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。他的教育计划旨在培养通过分配在核心化学工程课程中的一系列短模块，对化学工程课程进行分子层面的洞察。这里提出的研究的分子建模和实验结果将集成到模块中，这些模块旨在根据分子参数预测热导率、粘度和汽液平衡等基本工程特性。他提出的另一项教育工作是为期两周的胶体/界面工程迷你课程，强调生物技术和胶体科学之间的联系。