Bright individuals: Applications of fluorescent protein-based reporter systems in single-cell cellular microbiology.

IF 2.6 2区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Microbiology Pub Date : 2024-03-01 Epub Date: 2024-01-18 DOI:10.1111/mmi.15227
Marc Schulte, Luisa Grotheer, Michael Hensel
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Abstract

Activation and function of virulence functions of bacterial pathogens are highly dynamic in time and space, and can show considerable heterogeneity between individual cells in pathogen populations. To investigate the complex events in host-pathogen interactions, single cell analyses are required. Fluorescent proteins (FPs) are excellent tools to follow the fate of individual bacterial cells during infection, and can also be deployed to use the pathogen as a sensor for its specific environment in host cells or host organisms. This Resources describes design and applications of dual fluorescence reporters (DFR) in cellular microbiology. DFR feature constitutively expressed FPs for detection of bacterial cells, and FPs expressed by an environmentally regulated promoter for interrogation of niche-specific cues or nutritional parameters. Variations of the basic design allow the generation of DFR that can be used to analyze, on single cell level, bacterial proliferation during infection, subcellular localization of intracellular bacteria, stress response, or persister state. We describe basic considerations for DFR design and review recent applications of DFR in cellular microbiology.

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明亮的个体:基于荧光蛋白的报告系统在单细胞微生物学中的应用。
细菌病原体毒力功能的激活和作用在时间和空间上都是高度动态的,病原体群体中的单个细胞之间也会表现出相当大的异质性。要研究宿主-病原体相互作用中的复杂事件,需要进行单细胞分析。荧光蛋白(FPs)是跟踪感染过程中单个细菌细胞命运的绝佳工具,也可将病原体用作宿主细胞或宿主生物体内特定环境的传感器。本资料介绍了细胞微生物学中双荧光报告基因(DFR)的设计和应用。双荧光报告器的特点是组成型表达的 FPs 用于检测细菌细胞,而由环境调控启动子表达的 FPs 则用于检测特定生态位线索或营养参数。通过对基本设计进行变异,可以生成单细胞水平的 DFR,用于分析感染过程中的细菌增殖、细胞内细菌的亚细胞定位、应激反应或宿主状态。我们将介绍 DFR 设计的基本考虑因素,并回顾 DFR 在细胞微生物学中的最新应用。
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来源期刊
Molecular Microbiology
Molecular Microbiology 生物-生化与分子生物学
CiteScore
7.20
自引率
5.60%
发文量
132
审稿时长
1.7 months
期刊介绍: Molecular Microbiology, the leading primary journal in the microbial sciences, publishes molecular studies of Bacteria, Archaea, eukaryotic microorganisms, and their viruses. Research papers should lead to a deeper understanding of the molecular principles underlying basic physiological processes or mechanisms. Appropriate topics include gene expression and regulation, pathogenicity and virulence, physiology and metabolism, synthesis of macromolecules (proteins, nucleic acids, lipids, polysaccharides, etc), cell biology and subcellular organization, membrane biogenesis and function, traffic and transport, cell-cell communication and signalling pathways, evolution and gene transfer. Articles focused on host responses (cellular or immunological) to pathogens or on microbial ecology should be directed to our sister journals Cellular Microbiology and Environmental Microbiology, respectively.
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