Protocols in Current Issue
    Extraction and Quantification of Polyphosphate (polyP) from Gram-negative Bacteria
    Authors:  Jan-Ulrik Dahl, Lihan Xie and Ursula Jakob, date: 09/20/2018, view: 1789, Q&A: 0
    [Abstract] Polyphosphate (polyP), a universally conserved biomolecule, is composed of up to 1,000 phosphate monomers linked via phosphoanhydride bonds. Reaching levels in bacteria that are in the high nmoles per mg protein range, polyP plays important roles in biofilm formation and colonization, general stress protection and virulence. Various protocols for ...
    Bacterial Competition Assay Based on Extracellular D-amino Acid Production
    Authors:  Laura Alvarez and Felipe Cava, date: 04/05/2018, view: 3283, Q&A: 0
    [Abstract] Bacteria live in polymicrobial communities under tough competition. To persist in a specific niche many species produce toxic extracellular effectors as a strategy to interfere with the growth of nearby microbes. One of such effectors are the non-canonical D-amino acids. Here we describe a method to test the effect of D-amino acid production in ...
    Bacterial Aggregation Assay in the Presence of Cyclic Lipopeptides
    Authors:  Pengyuan Xiu, Rui Liu, Dechao Zhang and Chaomin Sun, date: 01/05/2018, view: 3182, Q&A: 0
    [Abstract] Lipopeptides is an important class of biosurfactants having antimicrobial and anti-adhesive activity against pathogenic bacteria. These include surfactin, fengycin, iturin, bacillomycin, mycosubtilin, lichenysin, and pumilacidin (Arima et al., 1968; Naruse et al., 1990; Yakimov et al., 1995; Steller and Vater, 2000; ...
    Protein Expression Protocol for an Adenylate Cyclase Anchored by a Vibrio Quorum Sensing Receptor
    Authors:  Stephanie Beltz and Joachim E. Schultz, date: 01/20/2017, view: 4781, Q&A: 0
    [Abstract] The direct regulation of a mycobacterial adenylate cyclase (Rv1625c) via exchange of its membrane anchor by the quorum sensing receptor CqsS (Vibrio harveyi) has recently been reported (Beltz et al., 2016). This protocol describes the expression and membrane preparation for these chimeric proteins.
    A Bioassay Protocol for Quorum Sensing Studies Using Vibrio campbellii
    Authors:  Alberto J. Martín-Rodríguez and José J. Fernández, date: 07/20/2016, view: 6290, Q&A: 0
    [Abstract] Quorum Sensing (QS), or bacterial cell-to-cell communication, is a finely-tuned mechanism that regulates gene expression on a population density-dependent manner through the production, secretion and reception of extracellular signaling molecules termed autoinducers (AIs). Given that QS plays an important role in bacterial biofilm formation and ...
    [14C] Linoleic Acid Uptake and Fractionation Assay in Vibrio cholerae
    Authors:  Sarah C. Plecha and Jeffrey H. Withey, date: 12/20/2015, view: 4212, 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 ...
    A Filter Binding Assay to Quantify the Association of Cyclic di-GMP to Proteins
    Authors:  Disha Srivastava and Christopher M. Waters, date: 02/05/2015, view: 7066, Q&A: 0
    [Abstract] Cyclic di-GMP (c-di-GMP) is a ubiquitous second messenger that regulates many processes in bacteria including biofilm formation, motility, and virulence (Hengge, 2009). Analysis of c-di-GMP binding properties of bacterial proteins is an important step to characterize c-di-GMP signaling pathways. C-di-GMP binds numerous proteins such as ...
    Infant Rabbit Colonization Competition Assays
    Authors:  Yang Fu and John J. Mekalanos, date: 06/05/2014, view: 6712, Q&A: 0
    [Abstract] Enteric pathogenic bacteria such as Vibrio cholerae and enteropathogenic Escherichia coli (E. coli) cause life-threatening diarrheal diseases that have afflicted humans for centuries. Understanding the effectors required for intestinal colonization is very important to research on bacteria pathogenesis, and is also ...
    Assay for Adherence of Vibrio cholerae to Eukaryotic Cell Lines
    Authors:  Amit K. Dey, Abha Bhagat and Rukhsana Chowdhury, date: 04/20/2014, view: 7595, Q&A: 0
    [Abstract] Vibrio cholerae (V. cholerae) colonizes the intestinal epithelium and secretes cholera toxin (CT), a potent enterotoxin that causes severe fluid loss characteristic of the disease cholera. V. cholerae is a non-invasive Gram-negative bacterium that adheres to intestinal cells as well as a variety of different cell types. ...
    Measurement of Haemolysin Activities in Vibrio vulnificus
    Authors:  Hyun-Jung Lee, Jeong-A Kim, Mi-Ae Lee, Soon-Jung Park and Kyu-Ho Lee, date: 03/05/2014, view: 5323, Q&A: 0
    [Abstract] VvhA produced by Vibrio vulnificus exhibits cytolytic activity to human cells including erythrocytes. Since haemolysis by VvhA may provide iron for bacterial growth and pathogenicity, we investigated the expression of VvhA to elucidate the regulatory roles of Fur, a major transcription factor controlling iron-homeostasis. Fur repressed ...

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