RUI: Phage-Based Components for Nanoscale Assembly

ID：MPS/dmr/bmat(7623)0804944 PI：Johnson，Karl ORG：Hiverford College标题：RUI：基于噬菌体的组件用于纳米级组装INTELLECTUCTUAL优点：此建议旨在开发丝状细菌病毒M13作为组装高阶结构的组件。为了实现这一目标，PI提出了以下具体任务：(1)系统地探索与噬菌体蛋白外壳相连的单色荧光染料的荧光浓度、亮度和猝灭之间的关系，以优化对单个噬菌体进行成像和跟踪的能力。(2)利用化学交联法将FRET(Forster Resonance Energy Transfer)染料对和其他官能团连接起来，噬菌体研究沿延伸的蛋白质-DNA骨架的能量吸收和转移，为涉及光子能量吸收和转移的应用奠定基础。(3)开发一系列专门与镍、塑料、硅或半导体材料等不同底物结合的噬菌体集合，以便将噬菌体自组装单层(自组装单分子膜)靶向独特的表面。(4)开发利用生物分子亲和力相互作用连接噬菌体的方法，例如生物素-链霉亲和素结合或多肽卷曲形成，将噬菌体彼此连接并以指定的几何形状围绕枢纽连接，以便在这些SAM？基础上建立。(5)用荧光显微镜、原子力显微镜和电子显微镜研究羊毛状和索状噬菌体网络的结构基础。(6)将这些聚合测量扩展到使用数字荧光显微镜测量单个噬菌体光谱强度，以验证使用单个pH敏感的染料标记的相粒子(噬菌体)作为工具来测量超细空间尺度上的PH值，并将这些传感器整合到我们的更高阶集合中。BROADER影响：这个RUI项目将在哈弗福德学院进行，这是一所小型文理学院，拥有多样化的学生群体和本科生参与研究的强大传统。文科环境是国家-S未来科学和医学领袖的丰富训练场，该项目提供的机会将使各个层次的学生接触到研究过程。通过校外讲座和校内讲习班，中学生及其教师参加的外联活动将放大这些影响，并将通过对这一跨学科项目的支持，加强学院丰富的研究环境。每年将有4-6名学生参与实现这项提案的目标。研究团队将包括本科生和高年级学生，他们将在暑期和学期中进行科学研究。这一提议的培训方面通过在学生职业生涯的关键时刻帮助他们学习新事物并就未来方向做出决定，从而发展了美国科学基础设施中的人的部分。这些学生将通过参与这些实验，在全国会议上展示他们的工作，并在主要期刊上发表描述他们对这一共同努力的贡献的出版物，成为这个项目的完全参与者。在上一次瑞安给国际学生联合会的资助中，共有30名学生参加，其中70%的本科生是女性，43%是代表不足的团体的成员。目前的项目希望继续这些努力，以扩大对国家科学事业的参与。

ID: MPS/DMR/BMAT(7623) 0804944 PI: Johnson, Karl ORG: Haverford CollegeTitle: RUI: Phage-Based Components for Nanoscale AssemblyINTELLECTUAL MERIT: This proposal aims to develop the filamentous bacterial virus M13 as a component for the assembly of higher-order structures. To accomplish this the PI proposes the following specific tasks: (1) Systematically explore relationships between fluorochrome density, brightness, and quenching for single-color fluorochromes linked to the phage protein coat in order to optimize the ability to image and track individual phage. (2) Use cross-linking chemistries to link FRET (Forster Resonance Energy Transfer) dye pairs and other functional groups to phage and study energy absorption and transfer along the extended protein-DNA backbone in order to lay the groundwork for applications involving photonic energy absorption and transfer. (3) Develop a collection of phage that bind specifically to diverse substrates such as nickel, plastics, silicon, or semiconductor materials in order to target phage SAMs (self assembled monolayers) to unique surfaces. (4) Develop ways to join phage using biomolecular affinity interactions, such as biotin-streptavidin binding or peptide coiled-coil formation, to link phage to each other and around hubs in defined geometries in order to build on these SAM ?foundations?. (5) Study the structural bases of wool-like and cable-like phage networks using fluorescence, atomic force, and electron microscopies. (6) Extend these aggregate measurements to the measurement of individual phage spectral intensities using digital fluorescence microscopy to validate the use of individual pH-sensitive-dye-labeled phase particles (pHages) as tools for the measurement of pH values on ultrafine spatial scales and incorporate these sensors into our higher order assemblies.BROADER IMPACTS: This RUI project will take place at Haverford College, a small liberal arts institution with a diverse student body and a strong tradition of undergraduate student involvement in research. The liberal arts environment is a rich training ground for the nation?s future leaders in science and medicine, and the opportunities made possible by this project will expose students at all levels to the research process. These effects will be magnified by outreach activities involving secondary school students and their teachers through both off-campus talks and on-campus workshops, and will enhance the research-rich environment at the College through support of this interdisciplinary project. Between 4-6 students per year will be involved in accomplishing the objectives of this proposal. The research team will include both underclass students and seniors in the process of doing science over the summer and during academic terms. The training aspects of this proposal develop the human component of US scientific infrastructure by reaching students at a critical point in their careers where they are learning new things and making decisions about future directions. These students will be full participants in this project by their involvement in these experiments, their presentation of their work at national meetings, and their co-authorship on publications in leading journals describing their contributions to this shared endeavor. In a previous RUI grant to the PI involving a total of 30 students, 70% of the undergraduate trainees were female and 43% were members of underrepresented groups. The current project expects to continue these efforts to broaden participation in the nation's scientific enterprise.