Microbiology

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    Protocols in Current Issue
    Measurements of Root Colonized Bacteria Species
    Authors:  Vílchez Juan Ignacio, Yu Yang, Dian Yi and Huiming Zhang, date: 04/05/2021, view: 259, Q&A: 0
    [Abstract]

    Root-associated bacteria are able to influence plant fitness and vigor. A key step in understanding the belowground plant-bacteria interactions is to quantify root colonization by the bacteria of interest. Probably, genetic engineering with fluorescence markers is the most powerful way to monitor bacterial strains in plant. However, this could

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    Brain-localized and Intravenous Microinjections in the Larval Zebrafish to Assess Innate Immune Response
    Authors:  Alison M. Rojas and Celia E. Shiau, date: 04/05/2021, view: 302, Q&A: 0
    [Abstract]

    Creating a robust and controlled infection model is imperative for studying the innate immune response. Leveraging the particular strengths of the zebrafish model system, such as optical transparency, ex utero development, and large clutch size, allows for the development of methods that yield consistent and reproducible results. We created a

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    A Spectrofluorophotometrical Method Based on Fura-2-AM Probe to Determine Cytosolic Ca2+ Level in Pseudomonas syringae Complex Bacterial Cells
    [Abstract]

    Calcium signaling is an emerging mechanism by which bacteria respond to environmental cues. To measure the intracellular free-calcium concentration in bacterial cells, [Ca2+]i, a simple spectrofluorometric method based on the chemical probe Fura 2-acetoxy methyl ester (Fura 2-AM) is here presented using Pseudomonad bacterial cells. This is an

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    Assay for Assessing Mucin Binding to Bacteria and Bacterial Proteins
    [Abstract]

    Legionella pneumophila, a Gram-negative bacterium and the causative agent of Legionnaires’ disease, exports over 300 effector proteins/virulence factors, through its type II (T2SS) and type IV secretion systems (T4SS). One such T2SS virulence factor, ChiA, not only functions as a chitinase, but also as a novel mucinase, which we believe aids

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    Defined Mutant Library Sequencing (DML-Seq) for Identification of Conditional Essential Genes
    Authors:  Shuai Shao, Lifan Wei, Feng Xia, Yuanxing Zhang and Qiyao Wang, date: 03/05/2021, view: 428, Q&A: 0
    [Abstract]

    Transposon insertion sequencing (TIS) is an emerging technique which utilizes a massive transposon mutant library to screen specific phenotype and determine the conditional essential genetic requirements for bacterial fitness under distinct conditions combined with high-throughput parallel sequencing technology. Compared with a massive mutant

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    Plant ARGONAUTE Protein Immunopurification for Pathogen Cross Kingdom Small RNA Analysis
    Authors:  Florian Dunker, Bernhard Lederer and Arne Weiberg, date: 02/05/2021, view: 688, Q&A: 0
    [Abstract]

    Over the last decade, it has been noticed that microbial pathogens and pests deliver small RNA (sRNA) effectors into their host plants to manipulate plant physiology and immunity for infection, known as cross kingdom RNA interference. In this process, fungal and oomycete parasite sRNAs hijack the plant ARGONAUTE (AGO)/RNA-induced silencing complex

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    Preparation of Nippostrongylus brasiliensis Larvae for the Study of Host Skin Response
    Authors:  Tiffany Bouchery, Gillian Coakley and Nicola L. Harris, date: 12/20/2020, view: 610, Q&A: 0
    [Abstract]

    Hookworms are skin penetrating parasites, however in the laboratory the hookworm model Nippostrongylus brasiliensis, the parasite is traditionally administered subcutaneously bypassing the skin (epidermis and dermis). Here, we describe two complementary approaches for infecting mice with N. brasiliensis in order to study the skin immune responses.

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    Pea Aphid Rearing, Bacterial Infection and Hemocyte Phagocytosis Assay
    Authors:  Li Ma, Lu Liu and Zhiqiang Lu, date: 12/20/2020, view: 719, Q&A: 0
    [Abstract]

    Insects rely on the simple but effective innate immune system to combat infection. Cellular and humoral responses are interconnected and synergistic in insects’ innate immune system. Phagocytosis is one major cellular response. It is difficult to collect clean hemolymph from the small insect like pea aphid. Here, we provide a practicable

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    Generating Three-dimensional Human Granulomas in vitro to Study Mycobacterium tuberculosis-host Interaction
    Authors:  Ainhoa Arbués, Michael Kammüller and Damien Portevin, date: 11/20/2020, view: 961, Q&A: 0
    [Abstract] Granulomas are organized multicellular structures that constitute the hallmark of an infection by the human pathogen Mycobacterium tuberculosis (Mtb). A better understanding of the complex host-Mtb interactions within the granuloma’s environment may lead to new therapeutic or preventive tools to improve the control of the ...
    Giant Mimiviridae CsCl Purification Protocol
    Authors:  Lionel Bertaux, Audrey Lartigue and Sandra Jeudy, date: 11/20/2020, view: 753, Q&A: 0
    [Abstract] While different giant viruses’ purification protocols are available, they are not fully described and they use sucrose gradient that does not reach an equilibrium. Here, we report a protocol for the purification of members of the Mimiviridae family virions resulting from Acanthamoeaba castellanii infections. Viruses are ...



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