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

PI建议进行实验，以确定ATPS聚合物和操作条件，以便标记的DNA将强烈分配到一个相中，将极性蛋白质和未标记的DNA留在另一个相中。通过适当的分子设计，当体系加热到PNA-DNA识别络合物的熔融转变温度(Tm)以上时，PNA两亲性将从DNA产品中解离。他还将介绍一系列理论和实验技术，这些技术将被用来设计PNA两亲分子，使它们与质粒DNA特异结合，但TM处于ATP的有用范围内。除了开发一种有用的生化过程外，这里提出的PNA两亲性DNA结合络合物的分子水平设计也应该阐明分子间作用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布置一系列简短的模块来培养化学工程课程中分子水平的洞察力。这里提出的研究的分子建模和实验结果将被整合到模块中，这些模块旨在根据分子参数预测基本的工程性质，如导热系数、粘度和汽液平衡。他提出的另一个教育努力是为期两周的胶体/界面工程课程的迷你课程，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.