Protocols in Current Issue
    Detection of in vivo Protein Interactions in All Bacterial Compartments by Förster Resonance Energy Transfer with the Superfolder mTurquoise2 ox-mNeongreen FRET Pair
    Authors:  Nils Y. Meiresonne, Elisa Consoli, Laureen M.Y. Mertens and Tanneke den Blaauwen, date: 12/05/2019, view: 1551, Q&A: 0
    [Abstract] This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in all compartments of Escherichia coli by Förster Resonance Energy Transfer (FRET) using the Superfolder mTurquoise2 ox-mNeonGreen FRET pair (sfTq2ox-mNG). This FRET pair has more than twice the detection range for FRET ...
    Precision Technique for Splenectomy Limits Mouse Stress Responses for Accurate and Realistic Measurements for Investigating Inflammation and Immunity
    Authors:  Shengwen Calvin Li, Anthony D. Rangel and Mustafa H. Kabeer, date: 08/05/2019, view: 1969, Q&A: 0
    [Abstract] Splenectomy in an animal model requires a standardized technique utilizing best practice to avoid variability which can result in adverse impact to the animal resulting in flawed physiologic responses simply due to technique rather than to the studied variables. In the case of the spleen, often investigators are analyzing the animal immune or ...
    Detection of Protein Interactions in the Cytoplasm and Periplasm of Escherichia coli by Förster Resonance Energy Transfer
    Authors:  Nils Y. Meiresonne, Svetlana Alexeeva, René van der Ploeg and Tanneke den Blaauwen, date: 01/20/2018, view: 6063, Q&A: 0
    [Abstract] This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in Escherichia coli by Förster Resonance Energy Transfer (FRET). The described assay allows for the previously impossible in vivo screening of periplasmic protein-protein interactions. In FRET, excitation of a donor fluorescent ...
    Quantification of Densities of Bacterial Endosymbionts of Insects by Real-time PCR
    Author:  Daisuke Kageyama, date: 10/05/2017, view: 6485, Q&A: 0
    [Abstract] Increased attention has been paid to the endosymbiotic bacteria of insects. Because most insect endosymbionts are uncultivable, quantitative PCR (qPCR) is a practical and convenient method to quantify endosymbiont titers. Here we report a protocol for real-time qPCR based on SYBR Green I fluorescence as well as some tips to prevent possible ...
    Escherichia coli Infection of Drosophila
    Authors:  Charles Tracy and Helmut Krämer, date: 05/05/2017, view: 5666, Q&A: 0
    [Abstract] Following septic insults, healthy insects, just like vertebrates, mount a complex immune response to contain and destroy pathogens. The failure to efficiently clear bacterial infections in immuno-compromised fly mutants leads to higher mortality rates which provide a powerful indicator for genes with important roles in innate immunity. The ...
    CRISPR/Cas9 Editing of the Bacillus subtilis Genome
    Authors:  Peter E. Burby and Lyle A. Simmons, date: 04/20/2017, view: 13092, Q&A: 4
    [Abstract] A fundamental procedure for most modern biologists is the genetic manipulation of the organism under study. Although many different methods for editing bacterial genomes have been used in laboratories for decades, the adaptation of CRISPR/Cas9 technology to bacterial genetics has allowed researchers to manipulate bacterial genomes with ...
    Analysis of the Virulence of Uropathogenic Escherichia coli Strain CFT073 in the Murine Urinary Tract
    Authors:  Anna Waldhuber, Manoj Puthia, Andreas Wieser, Catharina Svanborg and Thomas Miethke , date: 02/05/2017, view: 7134, Q&A: 0
    [Abstract] This urinary tract infection model was used to monitor the efficacy of a new virulence factor of the uropathogenic Escherichia coli strain CFT073 in vivo. The new virulence factor which we designated TIR-containing protein C (TcpC) blocks Toll-like receptor signaling and the NLRP3 inflammasome signaling cascade by interacting ...
    Preparation of Purified Gram-positive Bacterial Cell Wall and Detection in Placenta and Fetal Tissues
    Authors:  Beth Mann, Lip Nam Loh, Geli Gao and Elaine Tuomanen, date: 12/05/2016, view: 6812, Q&A: 0
    [Abstract] Cell wall is a complex biopolymer on the surface of all Gram-positive bacteria. During infection, cell wall is recognized by the innate immune receptor Toll-like receptor 2 causing intense inflammation and tissue damage. In animal models, cell wall traffics from the blood stream to many organs in the body, including brain, heart, placenta and ...
    Evaluation of Nodulation Speed by Sinorhizobium Strains
    Authors:  Matthew S. Nelson, Chan Lan Chun and Michael J. Sadowksy, date: 08/05/2015, view: 6347, Q&A: 0
    [Abstract] Rhizobia interact symbiotically with legumes to form root nodules, where by rhizobia fix atmospheric dinitrogen into ammonia in exchange for carbon produced via photosynthesis. The symbiotic interaction is agriculturally important by reducing the need for fertilizer containing nitrogen. The root and stem nodule bacteria commonly include bacteria ...

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