Microbiology

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    Protocols in Current Issue
    Intracellular Invasion and Killing Assay to Investigate the Effects of Binge Alcohol Toxicity in Murine Alveolar Macrophages
    Authors:  Victor Jimenez Jr and Fernando P Monroy, date: 01/20/2019, view: 986, Q&A: 0
    [Abstract] Alcohol consumption has diverse and well-documented effects on the human immune system and its ability to defend against infective agents. While pulmonary related infections can occur in healthy humans, binge alcohol use is recognized as a major health risk factor (Nelson et al., 1991). Although binge alcohol consumption has been ...
    High-throughput Microscopic Analysis of Salmonella Invasion of Host Cells
    Authors:  Jakub Voznica, Jost Enninga and Virginie Stévenin, date: 09/20/2018, view: 1393, Q&A: 0
    [Abstract] Salmonella is a Gram-negative bacterium causing a gastro-enteric disease called salmonellosis. During the first phase of infection, Salmonella uses its flagella to swim near the surface of the epithelial cells and to target specific site of infection. In order to study the selection criteria that determine which host cells are ...
    Liposome Flotation Assay for Studying Interactions Between Rubella Virus Particles and Lipid Membranes
    Authors:  Kyoko Saito, Noriyuki Otsuki, Makoto Takeda and Kentaro Hanada, date: 08/20/2018, view: 2140, Q&A: 0
    [Abstract] Rubella virus (RuV) is an enveloped, positive-sense single-stranded RNA virus that is pathogenic to humans. RuV binds to the target cell via the viral envelope protein E1, but the specific receptor molecules on the target cell are yet to be fully elucidated. Here, we describe a protocol for liposome flotation assay to study direct interactions ...
    Sleeping Beauty Transposon-based System for Rapid Generation of HBV-replicating Stable Cell Lines
    Authors:  Jin-Wei Zheng, Jia-Li Cao and Quan Yuan , date: 07/05/2018, view: 2124, Q&A: 0
    [Abstract] The stable HBV-transfected cell lines, which based on stable integration of replication-competent HBV genome into hepatic cells, are widely used in basic research and antiviral drug evaluation against HBV. However, previous reported strategies to generate HBV-replicating cell lines, which primarily rely on random integration of exogenous DNA by ...
    In-vitro and in-planta Botrytis cinerea Inoculation Assays for Tomato
    Authors:  Jiajie Lian, Hongyu Han, Jiuhai Zhao and Chuanyou Li, date: 04/20/2018, view: 3703, Q&A: 0
    [Abstract] Botrytis cinerea (B. cinerea) attacks many crops of economic importance, represents one of the most extensively studied necrotrophic pathogens. Inoculation of B. cinerea and phenotypic analysis of plant resistance are key procedures to investigate the mechanism of plant immunity. Here we describe a protocol for B. ...
    Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells
    Authors:  Jorge Luis Ayala-Lujan and Fernando Ruiz-Perez, date: 04/20/2018, view: 2195, Q&A: 0
    [Abstract] Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory ...
    Quantification of Colibactin-associated Genotoxicity in HeLa Cells by In Cell Western (ICW) Using γ-H2AX as a Marker
    Authors:  Sophie Tronnet and Eric Oswald, date: 03/20/2018, view: 2367, Q&A: 0
    [Abstract] The genotoxin colibactin is produced by several species of Enterobacteriaceae. This genotoxin induces DNA damage, cell cycle arrest, senescence and death in eukaryotic cells (Nougayrède et al., 2006; Taieb et al., 2016). Here we describe a method to quantify the genotoxicity of bacteria producing colibactin following a ...
    Membrane Lipid Screen to Identify Molecular Targets of Biomolecules
    Authors:  John R. Jimah, Paul H. Schlesinger and Niraj H. Tolia, date: 08/05/2017, view: 4338, Q&A: 0
    [Abstract] Proteins that bind to and disrupt cell membranes may target specific phospholipids. Here we describe a protocol to identify the lipid targets of proteins and biomolecules. First, we describe a screen to identify lipids in membranes that are specifically bound by the biomolecule of interest. Second, we describe a method for determining if the ...
    Adhesion and Invasion Assay Procedure Using Caco-2 Cells for Listeria monocytogenes
    Authors:  Swetha Reddy and Frank Austin, date: 05/05/2017, view: 6767, Q&A: 0
    [Abstract] Listeria monocytogenes is an important Gram-positive foodborne pathogen that is a particular problem in ready-to-eat food. It has an ability to survive in harsh conditions like refrigeration temperatures and high salt concentrations and is known to cross intestinal, placental and blood-brain barriers. Several cancerous cell lines like ...
    Isolation of Exosomes from Semen for in vitro Uptake and HIV-1 Infection Assays
    Authors:  Marisa N. Madison, Jennifer L. Welch and Chioma M. Okeoma, date: 04/05/2017, view: 5505, Q&A: 0
    [Abstract] Exosomes are membranous extracellular nanovesicles of endocytic origin. Exosomes are known to carry host and pathogen-derived genomic, proteomic, lipidomic cargos and other extraneous molecules. Exosomes are secreted by diverse cell types into the extracellular milieu and are subsequently internalized by recipient neighboring or distal cells. Upon ...



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