EXP-SA: Explosives Tracking: A Microsystem for Detection of Bacterial Endospores as Self-Replicating Nucleic Acid Taggants

该提案提出了一个综合的研究和教育计划，旨在生产，检测和鉴定用于爆炸物追踪的细菌内生孢子标记物。 虽然这项研究的最直接应用与遏制生物恐怖分子的活动有关，但生物工程和传感器科学与工程的预期基本进展将对其他应用具有重大的社会意义，包括第一反应者活动，医疗保健，食品安全以及避免和减轻污染。研究人员建议将嗜热脂肪芽孢杆菌内生孢子的联合收割机生物工程与用于样品处理和标记物鉴定的微型装置相结合。表面声波（SAW）微滴混合/运输/培养系统将与分子挂锁探针技术相结合，用于生物工程内生孢子的灵敏鉴定。具体的研究任务是：（i）产生许多不同的芽孢杆菌孢子，每一个孢子都有一个独特的DNA序列或拼接到其基因组中的序列;（ii）研究和确定萌发孢子、裂解营养细菌、运输、混合和加热微滴样品所需的最佳SAW装置设计;（iii）设计用于DNA分离的子系统; ㈣开发一个用于DNA鉴定的荧光分子挂锁探针系统，该系统可与表面声波制造微系统平台一起有效运作; ㈤制造和测试拟议的原型鉴定系统。㈠通过缅因州大学（缅因州）正在进行的两个主要方案，即国家科学基金会教师研究经验方案和GK-12传感器方案，培训高中学生并与他们互动;（二）让来自缅因州和其他机构的本科生直接参与正在进行的NSF研究经验的保护伞下的研究项目（三）为涉及跨学科研究和设计项目的本科生确定适当的顶点项目;（四）通过将研究成果纳入每门课程，加强现有的研究生课程（1）微尺度生物工程和（2）声波器件的设计和制造; ㈤促进国家科学基金会跨学科多机构综合研究生教育和研究培训方案的功能基因组学，该方案涉及缅因大学、杰克逊实验室和缅因州医学中心研究所;（六）为硕士提供论文题目。和博士㈦在项目网站上以及通过会议和印刷品传播研究和教育材料。

This proposal presents an integrated research and educational plan directed toward the production, detection, and identification of bacterial endospore taggants for explosive tracking. While the most immediate application of the research is related to stemming the activities of bioterrorists, the anticipated fundamental advances in bioengineering and sensor science and engineering will have significant societal relevance to other applications, including first-responder activities, healthcare, food safety, and pollution avoidance and mitigation. Intellectual Merit The investigators propose to combine bioengineering of Bacillus stearothermophilus endospores with microdevices for sample processing and taggant identification. A surface acoustic wave (SAW) microdroplet mixing/transport/incubator system will be coupled with molecular padlock probe technology for sensitive identification of bioengineered endospores. The specific research tasks are to: (i) Generate a number of different Bacillus spores, each with a unique DNA sequence or sequences spliced into its genome; (ii) Investigate and identify the optimal SAW device designs needed to germinate spores, lyse vegetative bacteria, transport, mix, and heat microdroplet samples; (iii) Design subsystems for DNA isolation; (iv) Develop a fluorescence-based molecular padlock probe system for DNA identification that can operate effectively in conjunction with the SAW fabrication microsystem platform; (v) Fabricate and test the proposed prototype identification system. Broader Impacts Broader impacts will be achieved through the following programs and activities to: (i) Train and interact with high school audiences through two major ongoing programs at University of Maine (UMaine), NSF Research Experiences for Teachers (RET) and the GK-12 Sensors; (ii) Involve undergraduates from Maine and other institutions directly into the research project under the umbrella of the ongoing NSF Research Experience for Undergraduates (REU) program at the UMaine; (iii) Identify appropriate Capstone projects for undergraduates involving cross-disciplinary research and design projects; (iv) Enhance existing graduate level courses (1) Microscale Bioengineering and (2) Design and Fabrication of Acoustic Wave Devices by incorporating research results into each course; (v) Contribute to the interdisciplinary multi-institutional NSF Integrative Graduate Education and Research Traineeship (IGERT) program in functional genomics, which involves UMaine, the Jackson Laboratory, and the Maine Medical Center Research Institute; (vi) Provide thesis topics for M.S. and Ph.D. students; (vii) Disseminate the research and educational material on a project website, and through conferences and printed literature.