Prokaryotes

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    Plaque Assay to Determine Invasion and Intercellular Dissemination of Shigella flexneri in TC7 Human Intestinal Epithelial Cells
    Authors:  Atin Sharma and Andrea Puhar, date: 07/05/2019, view: 448, Q&A: 0
    [Abstract] Shigella flexneri invades the epithelial cells lining the gut lumen and replicates intracellularly. The specialized Type III Secretion System (T3SS) and its effector proteins, encoded on a large virulence plasmid, assist the bacterium to gain access to the cytosol. Thereafter Shigella disseminates to neighboring cells in an ...
    Gentamicin Protection Assay to Determine the Number of Intracellular Bacteria during Infection of Human TC7 Intestinal Epithelial Cells by Shigella flexneri
    Authors:  Atin Sharma and Andrea Puhar, date: 07/05/2019, view: 531, Q&A: 0
    [Abstract] Shigella flexneri is an intracellular bacterial pathogen that gains access to the gut epithelium using a specialized Type III Secretion System (T3SS). Various determinants mediating this invasive infection have been experimentally verified using the classical gentamicin protection assay presented here. In this assay epithelial cell lines ...
    Kinetic Characterization of the Shigella Type Three Secretion System ATPase Spa47 Using α-32P ATP
    Authors:  Heather B. Case and Nicholas E. Dickenson, date: 11/05/2018, view: 1258, Q&A: 0
    [Abstract] ATPases represent a diverse class of enzymes that utilize ATP hydrolysis to support critical biological functions such as driving ion pumps, providing mechanical work, unfolding/folding proteins, and supporting otherwise thermodynamically unfavorable chemical reactions. We have recently shown that the Shigella protein Spa47 is an ATPase ...
    Transfer of Large Contiguous DNA Fragments onto a Low Copy Plasmid or into the Bacterial Chromosome
    Authors:  Analise Z Reeves and Cammie F Lesser, date: 11/20/2016, view: 6042, Q&A: 0
    [Abstract] Bacterial pathogenicity islands and other contiguous operons can be difficult to clone using conventional methods due to their large size. Here we describe a robust 3-step method to transfer large defined fragments of DNA from virulence plasmids or cosmids onto smaller autonomously replicating plasmids or directly into defined sites in the ...
    Secretion Assay in Shigella flexneri
    Authors:  Jonathan Reinhardt and Michael Kolbe, date: 11/20/2014, view: 7131, Q&A: 0
    [Abstract] Shigella flexneri (S. flexneri) is a Gram-negative bacterium that causes gastroenteritis and shigellosis in humans. In order to establish and maintain an infection, S. flexneri utilises a type three secretion system (T3SS) to deliver virulence factors called effector proteins into the cytoplasm of host cells, ...
    Design of a Transcription-based Secretion Activity Reporter (TSAR) for the Type III Secretion Apparatus of Shigella flexneri and Uses Thereof
    Authors:  François Xavier Campbell-Valois, Pamela Schnupf and Philippe J. Sansonetti, date: 10/20/2014, view: 6872, Q&A: 1
    [Abstract] Many gram-negative bacterial pathogens, including Shigella flexneri, are able to translocate bacterial proteins, dubbed effectors, across the host cell plasma membrane into the host cell cytosol using a syringe-like structure, the type three secretion apparatus (T3SA). While some bacteria use their T3SA to modulate their phagosomal ...
    Detection of the Secreted and Cytoplasmic Fractions of IpaB, IpaC and IpaD by Lysozyme Permeabilization
    Authors:  François Xavier Campbell-Valois, Pamela Schnupf and Philippe J. Sansonetti, date: 10/20/2014, view: 6105, Q&A: 0
    [Abstract] Gram negative bacterial pathogens, such as Shigella flexneri, which possess a Type Three Secretion System (T3SS), are able to transfer bacterial proteins, dubbed translocators and effectors, from their cytoplasm into the cytoplasm of their host cells using a syringe like needle complex. For Shigella, it has been shown that during ...
    Shigella IpaD and IpaB Surface Localizations
    Authors:  Lionel Schiavolin, Alaeddine Meghraoui and Abdelmounnaim Allaoui, date: 11/20/2013, view: 6669, Q&A: 1
    [Abstract] Shigella uses a type III secretion system to invade host cell and to cause disease. Secretion control and insertion of a translocation pore into cell membrane are critical steps for pathogenesis and are tightly linked to the formation of the needle tip complex formed by the IpaB and IpaD proteins (Veenendaal et al., 2007). ...



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