Antibiotic resistance is an alarming public health concern that affects millions of individuals across the globe each year. A major challenge in the development of effective antibiotics lies in their limited ability to permeate cells, noting that numerous susceptible antibiotic targets reside within the bacterial cytosol. Consequently, improving the cellular permeability is often a key consideration during antibiotic development, underscoring the need for reliable methods to assess the permeability of molecules across cellular membranes. Currently, methods used to measure permeability often fail to discriminate between the arrival within the cytoplasm and the overall association of molecules with the cell. Additionally, these techniques typically possess throughput limitations. In this work, we describe a luciferase-based assay designed for assessing the permeability of molecules in the cytosolic compartment of Gram-negative bacteria. Our findings demonstrate a robust system that can elucidate the kinetics of intracellular antibiotic accumulation in live bacterial cells in real time.

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基于生物发光的大肠杆菌胞浆内抗生素蓄积测定

抗生素耐药性是一个令人担忧的公共卫生问题，每年影响全球数百万人。开发有效抗生素的一个主要挑战在于它们渗透细胞的能力有限，并注意到许多敏感的抗生素靶点存在于细菌细胞质中。因此，提高细胞通透性通常是抗生素开发过程中的一个关键考虑因素，这强调了需要可靠的方法来评估分子跨细胞膜的通透性。目前，用于测量渗透性的方法通常无法区分到达细胞质内和分子与细胞的整体关联。此外，这些技术通常具有吞吐量限制。在这项工作中，我们描述了一种基于荧光素酶的测定，旨在评估革兰氏阴性细菌胞质室中分子的渗透性。我们的研究结果证明了一个强大的系统，可以实时阐明活细菌细胞内抗生素积累的动力学。