Probing the electrochemical dynamics of germination in Bacillus Subtilis spores

项目概述：这个由MRC资助的博士培训伙伴关系（DTP）将尖端的分子和分析科学与数据分析中的创新计算方法结合在一起，使学生能够解决以假设为主导的生物医学研究问题。这是一个为期4年的计划，其第一年涉及一系列教学模块和两个基于实验室的研究项目，导致跨学科生物医学研究硕士学位。前两个学期包括一系列教学模块，让学生在多学科科学中获得坚实的基础。学生还参加了一系列由学术和行业专家领导的大师班，这些专家在分子，细胞和组织动力学，微生物学和感染，应用生物医学技术以及人工智能和数据科学领域。在第三和夏季学期，学生在他们选择的实验室进行两个为期11周的研究项目。项目：几种细菌在恶劣的条件下形成孢子，但当环境再次变得适合居住时，它们可以重新萌发。这些孢子具有广泛的用途，从益生菌到引起炭疽的生物恐怖剂，并且在食品工业中存在问题，因此是生物医学研究的重要课题。休眠孢子有一个坚韧的多层外壳，耐热，耐辐射，耐极端pH值和毒素，使它们能够以植物状态存活多年。环境中的营养素触发萌发，使孢子恢复为活性细菌。虽然已知有基因参与发芽，但尚不清楚它们如何感知这些刺激并激活级联机制。我们假设发芽是由电化学特征介导的，包括离子（例如钙、钠等），Asally实验室最近的一篇论文表明，孢子的形成是由带电粒子控制的。我们建议使用相同的技术来研究细菌枯草芽孢杆菌中萌发的电生理学，并通过扫描电化学探针显微镜（SEPM）提高定量。这是一个多功能成像平台，使用尖端直径为10-100 nm的玻璃探针，扫描样品并高精度测量离子流和氧化还原特性。这个项目将要求学生使用微生物学技术与孢子一起工作，以及调查与发芽有关的基因缺陷的突变体。学生将接受培训，使用各种SEPM和用于控制显微镜的软件，这需要修改硬件和软件以优化设置。此外，学生将使用统计软件包来分析数据和模拟软件包来解释SEPM数据。因此，该项目将涵盖MRC规定的定量和跨学科技能培训主题。

Programme overview:This MRC-funded doctoral training partnership (DTP) brings together cutting-edge molecular and analytical sciences with innovative computational approaches in data analysis to enable students to address hypothesis-led biomedical research questions. This is a 4-year programme whose first year involves a series of taught modules and two laboratory-based research projects that lead to an MSc in Interdisciplinary Biomedical Research. The first two terms consist of a selection of taught modules that allow students to gain a solid grounding in multidisciplinary science. Students also attend a series of masterclasses led by academic and industry experts in areas of molecular, cellular and tissue dynamics, microbiology and infection, applied biomedical technologies and artificial intelligence and data science. During the third and summer terms students conduct two eleven-week research projects in labs of their choice. Project:Several species of bacteria sporulate in harsh conditions but can germinate back to activity when their environment becomes habitable again. These spores have a wide variety of uses ranging from probiotics to bioterror agents causing anthrax, and are problematic to inactivate in the food industry, thus an important topic of biomedical research. The dormant spores have a tough multi-layered shell resistant to heat, radiation, extreme pH, and toxins allowing them to survive in a vegetative state for years. Nutrients in the environment trigger germination, which recovers the spores to active bacteria. Though there are genes known to be involved in germination, it is unknown how they can sense such stimuli and activate the cascade of mechanisms. We hypothesise that germination is mediated by electrochemical features, including ions (e.g. calcium, sodium, etc.), membrane voltage and pH. A recent paper from the Asally lab demonstrated that spore formation was quality controlled by charged particles. We propose to use the same techniques to investigate the electrophysiology of germination in the bacteria Bacillus subtilis, and improve the quantification by scanning electrochemical probe microscopy (SEPM). This is a multifunctional imaging platform that uses glass probes with a tip diameter of 10-100 nm which scans samples and measure the flow of ions and redox properties with high precision. This project will require the student to use microbiological techniques to work with the spores as well as investigate mutants with defects in genes involved in germination. The student will be trained to use various the SEPMs and the software used to control the microscopes which requires modifying hardware and software for optimising the setup. Additionally, the student will be using statistical packages to analyse data and simulation packages for interpreting SEPM data. Hence, this project will cover the quantitative and interdisciplinary skills training themes set out by the MRC.