Investigating the molecular determinants of inter-bacterial competition

许多细菌病原体使用VI型分泌系统（T6SS）纳米机器将有毒的“效应”蛋白直接发射到靶细胞中。越来越明显的是，T6SS在多种革兰氏阴性细菌（包括重要的人类病原体）的毒力和竞争力中起着关键作用。许多不同的细菌，包括重要的病原体，可以使用t6ss靶向其他细菌细胞，杀死或抑制竞争对手。因此，“抗菌”t6ss提供了一种竞争机制，允许病原体在多微生物感染部位或环境库中增殖，并最终导致疾病。重要的是，了解T6SS抗菌效应物对目标细菌细胞的致命后果，为发现杀死或抑制细菌病原体的新方法提供了潜力。在这个项目中，我们的目标是利用遗传、生化、生物物理和结构方法的结合，确定一种或多种新型抗菌效应物的毒性的分子基础。解释跨学科的界面：这个项目将发展我在生物科学-物理界面以及基础生物化学方面的跨学科知识和技能。这将通过新毒素的纯化来进行，这将使我们能够使用各种生化分析来评估它们的毒性活性。这些毒素也将用于结晶并随后解决其结构的尝试。项目是否需要大量的定量分析技能？3号。项目是否需要大量的整体生理技能？否

Many bacterial pathogens use the Type VI secretion system (T6SS) nanomachine to fire toxic 'effector' proteins directly into target cells. It is becoming increasingly apparent that the T6SS plays a key role in the virulence and competitiveness of diverse Gram-negative bacteria, including important human pathogens. Many, diverse bacteria, including important pathogens, can use T6SSs to target other bacterial cells, killing or inhibiting rivals. 'Anti-bacterial' T6SSs thus provide a competitive mechanism to allow pathogens to proliferate in polymicrobial infection sites or environmental reservoirs and ultimately cause disease. Importantly, understanding the lethal consequences of T6SS anti-bacterial effectors on targeted bacterial cells offers the potential to uncover new ways to kill or inhibit bacterial pathogens. In this project we aim to determine the molecular basis of the toxicity of one or more novel anti-bacterial effectors, using a combination of genetic, biochemical, biophysical and structural approaches.1.Explain interdisciplinary interface: This project will develop my interdisciplinary knowledge and skills at the interface of bioscience-physics, as well as fundamental biochemistry. This will be conducted through purification of novel toxins, which shall enable us to evaluate their toxic activity using various biochemical assays. These toxins will also be used in attempts to crystallize and subsequently solve their structures.2.Does project require significant amount of quantitative skills? NO 3.Does project require significant amount of whole organism physiology skills? NO