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

PI提出了鉴定ATPS聚合物和操作条件的实验，以便标记的DNA将强烈地分裂到一个相，留下极性蛋白质和未标记的DNA在另一个相。通过适当的分子设计，当加热系统高于PNA-DNA识别复合物的熔化转变温度（Tm）时，PNA两亲体将与DNA产物分离。他还将描述一系列的理论和实验技术，这些技术将用于设计PNA两亲体，使它们能够特异性地结合质粒DNA，但在ATPS的有用范围内具有Tm。除了开发有用的生化过程外，本文提出的PNA两亲体- 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结构和功能的贡献。他的教育计划旨在通过在核心化学工程课程中分配的一系列短模块，培养化学工程课程中分子水平的洞察力。这里提出的分子建模和实验研究结果将集成到模块中，这些模块旨在从分子参数预测基本的工程特性，如导热性，粘度和汽液平衡。他提出的另一项教育努力是一个为期两周的胶体/界面工程迷你课程，强调生物技术和胶体科学之间的联系。

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.