Microbiology

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    Protocols in Current Issue
    Cryptococcus neoformans Virulence Assay Using a Galleria mellonella Larvae Model System
    Authors:  Piotr R. Stempinski, Daniel F. Q. Smith and Arturo Casadevall, date: 08/05/2022, view: 383, Q&A: 0
    [Abstract]

    Cryptococcus neoformans is a human pathogenic fungus that can cause pulmonary infections and meningitis in both immunocompromised and otherwise healthy individuals. Limited treatment options and a high mortality rate underlie the necessity for extensive research of the virulence of C. neoformans. Here we describe a detailed protocol for using the

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    Cost Effective Method for gDNA Isolation from the Cecal Content and High Yield Procedure for RNA Isolation from the Colonic Tissue of Mice
    Authors:  Sohini Mukhopadhyay and Palok Aich, date: 08/05/2022, view: 216, Q&A: 0
    [Abstract]

    Microbiome studies are quickly gaining momentum. Since most of the resident microbes (consisting of bacteria, fungi, and viruses) are difficult to culture, sequencing the microbial genome is the method of choice to characterize them. It is therefore important to have efficient methodology for gDNA isolation of gut microbes. Mouse models are widely

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    VirScan: High-throughput Profiling of Antiviral Antibody Epitopes
    Authors:  Ellen L. Shrock, Christine L. Shrock and Stephen J. Elledge, date: 07/05/2022, view: 1090, Q&A: 0
    [Abstract]

    Profiling the specificities of antibodies can reveal a wealth of information about humoral immune responses and the antigens they target. Here, we present a protocol for VirScan, an application of the phage immunoprecipitation sequencing (PhIP-Seq) method for profiling the specificities of human antiviral antibodies. Accompanying this protocol is

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    A β-glucuronidase (GUS) Based Bacterial Competition Assay to Assess Fine Differences in Fitness during Plant Infection
    Authors:  Julien S. Luneau, Laurent D. Noël, Emmanuelle Lauber and Alice Boulanger, date: 07/05/2022, view: 886, Q&A: 0
    [Abstract]

    Competition assays are a simple phenotyping strategy that confront two bacterial strains to evaluate their relative fitness. Because they are more accurate than single-strain growth assays, competition assays can be used to highlight slight differences that would not otherwise be detectable. In the frame of host-pathogens interactions, they can be

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    Purification of the Bacterial Amyloid “Curli” from Salmonella enterica Serovar Typhimurium and Detection of Curli from Infected Host Tissues
    [Abstract]

    Microbiologists are learning to appreciate the importance of “functional amyloids” that are produced by numerous bacterial species and have impacts beyond the microbial world. These structures are used by bacteria to link together, presumably to increase survival, protect against harsh conditions, and perhaps to influence cell-cell

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    Ex vivo Human Skin Infection with Herpes Simplex Virus 1
    Authors:  Nydia C. De La Cruz, Maureen Möckel, Lisa Wirtz and Dagmar Knebel-Mörsdorf, date: 05/05/2022, view: 630, Q&A: 0
    [Abstract]

    Although herpes simplex virus 1 (HSV-1) is a well-studied virus, how the virus invades its human host via skin and mucosa to reach its receptors and initiate infection remains an open question. For studies of HSV-1 infection in skin, mice have been used as animal models. Murine skin infection can be induced after injection or scratching of the

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    Production of Recombinant Hepatitis B virus (HBV) and Detection of HBV in Infected Human Liver Organoids
    Authors:  Tanvir Hossain, Shahla Romal and Tokameh Mahmoudi, date: 04/20/2022, view: 751, Q&A: 0
    [Abstract]

    The absence of long term, primary untransformed in vitro models that support hepatitis B virus (HBV) infection and replication have hampered HBV pre-clinical research, which was reflected in the absence of a curative therapy until recently. One of the limitations for in vitro HBV research has been the absence of high titer and pure recombinant HBV

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    Generation, Maintenance and HBV Infection of Human Liver Organoids
    Authors:  Shahla Romal, Tanvir Hossain and Tokameh Mahmoudi, date: 03/20/2022, view: 1314, Q&A: 0
    [Abstract]

    Hepatitis B virus (HBV) infection represents a major public health problem infecting approximately 400 million people worldwide. Despite the availability of a preventive vaccine and anti-viral therapies, chronic HBV infection remains a major health issue because it increases the risk of developing liver cirrhosis and hepatocellular carcinoma

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    Three-dimensional Models of the Nasopharynx for the Study of Epstein-Barr Virus Infection
    [Abstract]

    The ubiquitous and cancer-associated Epstein-Barr virus (EBV) is associated with nearly all cases of nasopharyngeal carcinoma (NPC). Nasopharyngeal tissue is comprised of both pseudostratified and stratified epithelium, which are modeled in three-dimensional (3-D) cell culture. The cellular origin of EBV-associated NPC is as yet unknown, but both

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    Amber Suppression Technology for Mapping Site-specific Viral-host Protein Interactions in Mammalian Cells
    Authors:  Nur Firdaus Isa, Olivier Bensaude and Shona Murphy, date: 02/05/2022, view: 1482, Q&A: 0
    [Abstract]

    Probing the molecular interactions of viral-host protein complexes to understand pathogenicity is essential in modern virology to help the development of antiviral therapies. Common binding assays, such as co-immunoprecipitation or pull-downs, are helpful in investigating intricate viral-host proteins interactions. However, such assays may miss

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