Microbiology

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    Protocols in Current Issue
    A Yeast Chromatin-enriched Fractions Purification Approach, yChEFs, from Saccharomyces cerevisiae
    [Abstract] We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we bypass subcellular fractionation requirements involved when using ...
    Isolation of Outer Membrane Vesicles from Phytopathogenic Xanthomonas campestris pv. campestris
    Authors:  Gideon Mordukhovich and Ofir Bahar, date: 03/05/2017, view: 6397, Q&A: 3
    [Abstract] Gram-negative bacteria naturally release outer membrane vesicles (OMVs) to the surrounding environment. OMVs contribute to multiple processes, such as cell-cell communication, delivery of enzymes and toxins, resistance to environmental stresses and pathogenesis. Little is known about OMVs produced by plant-pathogenic bacteria, and their ...
    Preparation of Outer Membrane Vesicles from Myxococcus xanthus
    Authors:  James E. Berleman, Marcin Zemla, Jonathan P Remis and Manfred Auer, date: 01/20/2016, view: 7472, Q&A: 0
    [Abstract] Outer membrane vesicles (OMVs) represent a unique sub-cellular compartment of bacteria that may act as a scaffold for various extracellular activities, including intercellular signaling. Myxococcus xanthus (M. xanthus) is a predatory bacterium that engages in cell-cell behaviors such as fruiting body formation and contact ...
    [14C] Linoleic Acid Uptake and Fractionation Assay in Vibrio cholerae
    Authors:  Sarah C. Plecha and Jeffrey H. Withey, date: 12/20/2015, view: 4648, Q&A: 0
    [Abstract] The gram-negative curved bacillus Vibrio cholerae (V. cholerae) causes the severe diarrheal illness cholera. The work presented here is to assess whether unsaturated fatty acids (UFAs), such as linoleic acid, have the potential to directly affect proteins involved in DNA binding because they are able to enter the cell. In this ...
    Membrane Flotation Assay
    Authors:  Dorothee A Vogt and Melanie Ott, date: 04/05/2015, view: 9845, Q&A: 0
    [Abstract] Many postitive-stranded RNA viruses, such as Hepatitis C virus (HCV), highjack cellular membranes, including the Golgi, ER, mitchondria, lipid droplets, and utilize them for replication of their RNA genome or assembly of new virions. By investigating how viral proteins associate with cellular membranes we will better understand the roles of ...
    Rapid Preparation of Unsheathed Bacterial Flagella
    Authors:  Soazig Le Guyon, Mikael Rhen and Ute Römling, date: 03/20/2015, view: 7268, Q&A: 0
    [Abstract] The flagellum is required for bacterial swimming and swarming motility. In the biphasic Salmonella enterica serovar Typhimurium (S. Typhimurium), the flagellar filament is build up by two distinct monomeric subunits, flagellin FliC and FljB. S. Typhimurium has the ability to switch between two flagellins, FliC and FljB, ...
    Differentiation of Naturally Produced Extracellular Membrane Vesicles from Lipid Aggregation by Glucuronoxylomannan Immunogold Transmission Electron Microscopy in Bacillus subtilis
    Authors:  Lisa Brown, Geoff Perumal and Arturo Casadevall, date: 03/05/2015, view: 7064, Q&A: 0
    [Abstract] Recently, membrane vesicle (MV) production was described in Gram-positive bacteria, which harbor a variety of components such as toxins, antibiotic resistance proteins, proteases, DNA, and immune modulators. Free lipids have the ability to form micelles, thus it is important to rule out spontaneous association of lipids into vesicle-like ...
    Vesicle Isolation from Bacillus subtilis Biofilm
    Authors:  Lisa Brown, Anne Kessler and Arturo Casadevall, date: 03/05/2015, view: 8964, Q&A: 0
    [Abstract] Bacterial biofilms are associated clinically with many bacterial infections including those caused by bacteria such as Pseudomonas aeruginosa and Staphylococcus aureus. In recent years, extracellular vesicles produced by bacteria have been isolated from biofilm communities. Vesicles have been described in depth and can ...
    Fractionation by Ultracentrifugation of Gram Negative Cytoplasmic and Membrane Proteins
    Authors:  Sara M Sandrini, Richard Haigh and Primrose P. E. Freestone, date: 11/05/2014, view: 14809, Q&A: 1
    [Abstract] Protein fractionation is a useful separation process which divides membrane proteins (including those located in the outer and inner membrane) and cytoplasmic proteins into discrete fractions. Fractionation of proteins can simplify analysis of the numbers of proteins present, and therefore make easier to characterize any environmentally or ...
    Preparation of Outer Membrane Vesicle from Escherichia coli
    [Abstract] Outer membrane vesicles (OMVs) are spherical bilayered phospholipids of 20-200 nm in size produced from all Gram-negative bacteria and Gram-positive bacteria investigated to date. Previous biochemical and proteomic studies have revealed that the Gram-negative bacteria-derived OMVs are composed of various components like outer membrane proteins, ...



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