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Author:
Date:
2020-03-05
[Abstract] The emergence and rapid spread of multidrug resistance in bacteria have led to the urgent need for novel antibacterial agents. Membrane permeabilization is the mechanism for many antibacterial molecules that are being developed against gram-negative bacteria. Thus, to determine the efficacy of a potential antibacterial molecule, it is important to assess the change in bacterial membrane permeability after treatment. This study describes the protocol for the assays of outer and inner membrane permeability using the fluorescent probes N-phenyl-1-naphthylamine and propidium iodide. Compared with other experiments, such as electron microscopy and the assay of minimal bactericidal concentration, this methodology provides a simpler, faster, and cost-effective way of estimating the ...
[摘要] [摘要 ] 细菌中多药耐药性的出现和迅速扩散导致对新型抗菌剂的迫切需求。膜通透性是许多针对革兰氏阴性细菌的抗菌分子的机制。因此，为了确定潜在的抗菌分子的功效，重要的是评估治疗后细菌膜通透性的变化。这项研究描述了使用荧光探针N-苯基-1-萘胺和碘化丙啶测定外膜和内膜通透性的方案。与其他实验（例如电子显微镜和最小杀菌浓度的测定）相比，该方法提供了一种更简单，更快速且经济高效的方法来评估抗菌分子的膜通透性和杀菌功效。这项研究提出了一种相对于传统方案的优化方案，该方案是在与抗菌测定相同的培养条件下，将细菌与抗菌分子一起孵育，然后在不含肉汤和抗菌分子的缓冲液中检测荧光探针的信号。此协议可避免的效果养分缺乏对细菌的生理状态和抗菌molecul的干扰ES 朝向的F luorescent探针。因此，该方法可以有效，准确地评估膜的渗透性，并与从其他抗菌测定法（如最小抑菌浓度和时间杀灭曲线测定法）获得的结果相匹配。
[背景 ] ...
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