Protocols in Current Issue
    Measuring Homologous Recombination Rates between Chromosomal Locations in Salmonella
    Authors:  Gerrit Brandis, Sha Cao and Diarmaid Hughes, date: 02/05/2019, view: 1351, Q&A: 0
    [Abstract] Homologous recombination between two similar DNA molecules, plays an important role in the repair of double-stranded DNA breaks. Recombination can occur between two sister chromosomes, or between two locations of similar sequence identity within the same chromosome. The assay described here is designed to measure the rate of homologous ...
    High-throughput Microscopic Analysis of Salmonella Invasion of Host Cells
    Authors:  Jakub Voznica, Jost Enninga and Virginie Stévenin, date: 09/20/2018, view: 1774, Q&A: 0
    [Abstract] Salmonella is a Gram-negative bacterium causing a gastro-enteric disease called salmonellosis. During the first phase of infection, Salmonella uses its flagella to swim near the surface of the epithelial cells and to target specific site of infection. In order to study the selection criteria that determine which host cells are ...
    Extraction of Small Molecules from Fecal Samples and Testing of Their Activity on Microbial Physiology
    Authors:  Eduardo S. Alves, Rosana B. R. Ferreira and L. Caetano M. Antunes, date: 04/20/2018, view: 2592, Q&A: 0
    [Abstract] The human body is colonized by vast communities of microbes, collectively known as microbiota, or microbiome. Although microbes colonize every surface of our bodies that is exposed to the external environment, the biggest collection of microbes colonizing humans and other mammals can be found in the gastrointestinal tract. Given the fact that the ...
    Microbial Mutagenicity Assay: Ames Test
    Authors:  Urvashi Vijay, Sonal Gupta, Priyanka Mathur, Prashanth Suravajhala and Pradeep Bhatnagar, date: 03/20/2018, view: 14227, Q&A: 2
    [Abstract] The Microbial mutagenicity Ames test is a bacterial bioassay accomplished in vitro to evaluate the mutagenicity of various environmental carcinogens and toxins. While Ames test is used to identify the revert mutations which are present in strains, it can also be used to detect the mutagenicity of environmental samples such as drugs, dyes, ...
    Determination of Local pH Differences within Living Salmonella Cells by High-resolution pH Imaging Based on pH-sensitive GFP Derivative, pHluorin(M153R)
    Authors:  Yusuke V. Morimoto, Nobunori Kami-ike, Keiichi Namba and Tohru Minamino, date: 09/05/2017, view: 3308, Q&A: 0
    [Abstract] The bacterial flagellar type III protein export apparatus is composed of a transmembrane export gate complex and a cytoplasmic ATPase complex. The export apparatus requires ATP hydrolysis and the proton motive force across the cytoplasmic membrane to unfold and transport flagellar component proteins for the construction of the bacterial flagellum ...
    Evaluation of Plasmid Stability by Negative Selection in Gram-negative Bacteria
    Authors:  Damián Lobato Márquez and Laura Molina García, date: 05/05/2017, view: 6061, Q&A: 0
    [Abstract] Plasmid stability can be measured using antibiotic-resistance plasmid derivatives by positive selection. However, highly stable plasmids are below the sensitivity range of these assays. To solve this problem we describe a novel, highly sensitive method to measure plasmid stability based on the selection of plasmid-free cells following elimination ...
    Measurements of Free-swimming Speed of Motile Salmonella Cells in Liquid Media
    Authors:  Yusuke V. Morimoto, Keiichi Namba and Tohru Minamino, date: 01/05/2017, view: 4868, Q&A: 0
    [Abstract] Bacteria such as Escherichia coli and Salmonella enterica swim in liquid media using the bacterial flagella. The flagellum consists of the basal body (rotary motor), the hook (universal joint) and the filament (helical screw). Since mutants with a defect in flagellar assembly and function cannot swim smoothly, motility assay is ...
    Bacterial Intracellular Sodium Ion Measurement using CoroNa Green
    Authors:  Yusuke V. Morimoto, Keiichi Namba and Tohru Minamino, date: 01/05/2017, view: 5500, Q&A: 0
    [Abstract] The bacterial flagellar type III export apparatus consists of a cytoplasmic ATPase complex and a transmembrane export gate complex, which are powered by ATP and proton motive force (PMF) across the cytoplasmic membrane, respectively, and transports flagellar component proteins from the cytoplasm to the distal end of the growing flagellar structure ...
    In vitro Real-time Measurement of the Intra-bacterial Redox Potential
    Authors:  Joris van der Heijden and B. Brett Finlay, date: 09/05/2015, view: 5420, Q&A: 0
    [Abstract] All bacteria that live in oxygenated environments have to deal with oxidative stress caused by some form of exogenous or endogenous reactive oxygen species (ROS) (Imlay, 2013). Large quantities of ROS damage DNA, lipids and proteins which can eventually lead to bacterial cell death (Imlay, 2013). In contrast, smaller quantities of ROS can play ...
    Rapid Preparation of Unsheathed Bacterial Flagella
    Authors:  Soazig Le Guyon, Mikael Rhen and Ute Römling, date: 03/20/2015, view: 6660, 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, ...

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