Microbiology

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    Protocols in Current Issue
    Flow Cytometry-based Measurement of Reactive Oxygen Species in Cyanobacteria
    Authors:  Soumila Mondal and Shailendra P. Singh, date: 05/20/2022, view: 36, Q&A: 0

    Cyanobacteria are Gram-negative oxygen-producing photosynthetic bacteria that are useful in the pharmaceutical and biofuel industries. Monitoring of oxidative stress under fluctuating environmental conditions is important for determining the

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

    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,

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    Flow Cytometry-based Measurement of Reactive Oxygen Species in Cyanobacteria
    Authors:  Soumila Mondal and Shailendra P. Singh, date: 05/20/2022, view: 36, Q&A: 0
    [Abstract]

    Cyanobacteria are Gram-negative oxygen-producing photosynthetic bacteria that are useful in the pharmaceutical and biofuel industries. Monitoring of oxidative stress under fluctuating environmental conditions is important for determining the fitness, survival, and growth of cyanobacteria in the laboratory as well as in large scale cultivation

    ...
    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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    A Molecular Cloning and Sanger Sequencing-based Protocol for Detecting Site-specific DNA Methylation
    Authors:  Wei Guo, Anthony Cannon and Damon Lisch, date: 05/05/2022, view: 476, Q&A: 0
    [Abstract]

    DNA methylation is a conserved chemical modification, by which methyl groups are added to the cytosine of DNA molecules. Methylation can influence gene expression without changing the sequence of a particular gene. This epigenetic effect is an intriguing phenomenon that has puzzled biologists for years. By probing the temporal and spatial patterns

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    Expression, Purification, and in vitro Enzyme Activity Assay of a Recombinant Aldehyde Dehydrogenase from Thermus thermophilus, using an Escherichia coli host
    Authors:  Kim Shortall, Edmond Magner and Tewfik Soulimane, date: 05/05/2022, view: 551, Q&A: 0
    [Abstract]

    Based on previous in-depth characterisation, aldehyde dehydrogenases (ALDH) are a diverse superfamily of enzymes, in terms of both structure and function, present in all kingdoms of life. They catalyse the oxidation of an aldehyde to carboxylic acid using the cofactor nicotinamide adenine dinucleotide (phosphate) (NAD(P)+), and are often not

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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: 328, 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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    Bacterial Growth Curve Measurements with a Multimode Microplate Reader
    Authors:  Ariel T. Rogers, Kaitlin R. Bullard, Akash C. Dod and Yong Wang, date: 05/05/2022, view: 647, Q&A: 0
    [Abstract]

    Bacterial studies based on growth curves are common in microbiology and related fields. Compared to the standard photometer and cuvette based protocols, bacterial growth curve measurements with microplate readers provide better temporal resolution, higher efficiency, and are less laborious, while analysis and interpretation of the microplate-based

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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: 403, 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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    Tracking the Reversed Oxidative Tricarboxylic Acid Cycle in Bacteria
    [Abstract]

    Different pathways for autotrophic CO2 fixation can be recognized by the presence of genes for their specific key enzymes. On this basis, (meta)genomic, (meta)transcriptomic, or (meta)proteomic analysis enables the identification of the role of an organism or a distinct pathway in primary production. However, the recently discovered variant of the

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    Phytophthora sojae Transformation Based on the CRISPR/Cas9 System
    Authors:  Jingting Cao, Min Qiu, Wenwu Ye and Yuanchao Wang, date: 03/20/2022, view: 532, Q&A: 0
    [Abstract]

    Phytophthora sojae is a model species for the study of plant pathogenic oomycetes. The initial research on gene function using Phytophthora was mainly based on gene silencing technology. Recently, the CRISPR/Cas9-mediated genome editing technology was successfully established in P. sojae and widely used in oomycetes. In this protocol, we describe

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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: 911, 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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