Search

Prokaryotes

Categories

    Protocols in Current Issue
    Snapshots of the Signaling Complex DesK:DesR in Different Functional States Using Rational Mutagenesis and X-ray Crystallography
    [Abstract] We have developed protocols to generate site-specific variants of the histidine-kinase DesK and its cognate response regulator DesR, conducive to trapping different signaling states of the proteins. Co-expression of both partners in E. coli, ensuring an excess of the regulator, was essential for soluble production of the DesK:DesR ...
    Analysis of N-acetylmuramic acid-6-phosphate (MurNAc-6P) Accumulation by HPLC-MS
    Authors:  Marina Borisova and Christoph Mayer, date: 08/05/2017, view: 420, Q&A: 0
    [Abstract] We describe here in detail a high-performance liquid chromatography-mass spectrometry (HPLC-MS)-based method to determine N-acetylmuramic acid-6-phosphate (MurNAc-6P) in bacterial cell extracts. The method can be applied to both Gram-negative and Gram-positive bacteria, and as an example we use Escherichia coli cells in this ...
    Determination of NO and CSF Levels Produced by Bacillus subtilis
    [Abstract] The cell-to-cell communication and division of labour that occurs inside a beneficial biofilm produce significant differences in gene expression compared with the gene expression pattern of cells grew under planktonic conditions. In this sense, the levels of NO (nitric oxide) and CSF (Competence Sporulation Stimulating Factor) produced in Bacillus ...
    Culturing Bacteria from Caenorhabditis elegans Gut to Assess Colonization Proficiency
    [Abstract] Determining an accurate count of intestinal bacteria from Caenorhabditis elegans is one critical way to assess colonization proficiency by a given bacteria. This can be accomplished by culturing appropriate dilutions of worm gut bacteria on selective or differential agarized media. Because of the high concentration of bacteria in gut ...
    CRISPR/Cas9 Editing of the Bacillus subtilis Genome
    Authors:  Peter E. Burby and Lyle A. Simmons, date: 04/20/2017, view: 1622, Q&A: 0
    [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 ...
    Measurement of the Galactanase Activity of the GanB Galactanase Protein from Bacillus subtilis
    Author:  Hildegard Watzlawick, date: 04/05/2017, view: 797, Q&A: 0
    [Abstract] The activity of the endo-β-1,4-galactanase GanB from B. subtilis on the high molecular weight β-1,4-galactan was determined quantitatively by the measurement of the increase of the reducing power or with the dyed substrate Azo-galactan. The generated degradation products were analyzed using thinlayer-chromatography (TLC) or ...
    Purification of 70S Ribosomes from Bacillus subtilis
    Authors:  Shota Suzuki, Genki Akanuma and Fujio Kawamura, date: 04/05/2015, view: 3318, Q&A: 0
    [Abstract] The eubacterial ribosome (70S) is a macromolecular complex that is composed of a small (30S) subunit and a large (50S) subunit. The small subunit comprises the 16S ribosomal RNA (rRNA) and more than 20 ribosomal proteins (r-proteins), whereas the large subunit comprises the 23S and 5S rRNAs and more than 30 r-proteins. Bacillus subtilis ( ...
    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: 3033, 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: 3650, 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 ...
    Isolation of the Secretome from Bacillus subtilis
    Authors:  Juri Niño Bach and Marc Bramkamp, date: 01/20/2014, view: 3654, Q&A: 0
    [Abstract] Bacteria are commonly known to secret proteins in large amounts into the surrounding environment in high concentrations via various pathways. These proteins can be involved in numerous processes like cell-cell communication, exopolymer formation but also metabolic active enzymes are secreted that are interesting for industrial production of ...