RUI: Phage-Based Components for Nanoscale Assembly

ID: MPS/DMR/BMAT(7623) 0804944 PI: Johnson， Karl ORG: Haverford college标题：RUI：基于噬菌体的纳米级组装组件知识优点：该建议旨在开发丝状细菌病毒M13作为高阶结构组装的组件。为了实现这一目标，PI提出了以下具体任务：(1)系统探索与噬菌体蛋白外壳连接的单色荧光染料的荧光密度、亮度和猝灭之间的关系，以优化对单个噬菌体的成像和跟踪能力。(2)利用交联化学将FRET （Forster Resonance Energy Transfer）染料对等功能基团与噬菌体连接，研究沿延伸蛋白- dna主干的能量吸收和转移，为涉及光子能量吸收和转移的应用奠定基础。(3)开发一组噬菌体，这些噬菌体可以特异性结合不同的底物，如镍、塑料、硅或半导体材料，以便将噬菌体sam（自组装单层）靶向到独特的表面。(4)开发利用生物分子亲和相互作用连接噬菌体的方法，如生物素-链亲和素结合或肽盘绕形成，将噬菌体彼此连接，并以确定的几何形状围绕中心连接，以建立这些SAM基础。(5)利用荧光、原子力和电子显微镜研究羊毛状和电缆状噬菌体网络的结构基础。(6)将这些集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，以验证单个pH敏感染料标记相颗粒（噬菌体）作为超细空间尺度上pH值测量工具的使用，并将这些传感器整合到我们的高阶组件中。更广泛的影响：这个RUI项目将在哈弗福德学院进行，哈弗福德学院是一个小型的文理学院，学生群体多样化，本科生参与研究的传统很强。文科环境是国家丰富的训练基地。美国未来在科学和医学领域的领导者，以及该项目所带来的机会，将使各级学生接触到研究过程。通过校外讲座和校内工作坊，让中学生和他们的老师参与到这些活动中，这些效果将会被放大，并通过支持这个跨学科项目，加强学院的研究环境。每年将有4-6名学生参与完成该提案的目标。研究小组将包括低年级学生和高年级学生，在夏季和学期期间进行科学研究。该提案的培训方面发展了美国科学基础设施的人力组成部分，帮助处于职业生涯关键阶段的学生学习新事物并对未来方向做出决定。这些学生将通过参与这些实验，在国家会议上展示他们的工作，并在主要期刊上共同撰写出版物来描述他们对这一共同努力的贡献，从而成为该项目的全面参与者。在之前的一项由RUI资助的项目中，共有30名学生参与，其中70%的本科生是女性，43%是代表性不足的群体成员。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.