Prokaryotes

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    Protocols in Current Issue
    Non-invasive Quantification of Cell Wall Porosity by Fluorescence Quenching Microscopy
    Authors:  Xiaohui Liu, Thomas Günther Pomorski and Johannes Liesche, date: 08/20/2019, view: 217, Q&A: 0
    [Abstract] All bacteria, fungi and plant cells are surrounded by a cell wall. This complex network of polysaccharides and glycoproteins provides mechanical support, defines cell shape, controls cell growth and influences the exchange of substances between the cell and its surroundings. Despite its name, the cell wall is a flexible, dynamic structure. ...
    Extraction and Quantification of Polyphosphate (polyP) from Gram-negative Bacteria
    Authors:  Jan-Ulrik Dahl, Lihan Xie and Ursula Jakob, date: 09/20/2018, view: 1781, 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 ...
    Implementation of Blue Light Switchable Bacterial Adhesion for Design of Biofilms
    Authors:  Fei Chen and Seraphine V. Wegner, date: 06/20/2018, view: 2232, Q&A: 0
    [Abstract] Control of bacterial adhesions to a substrate with high precision in space and time is important to form a well-defined biofilm. Here, we present a method to engineer bacteria such that they adhere specifically to substrates under blue light through the photoswitchable proteins nMag and pMag. This provides exquisite spatiotemporal remote control ...
    Quantification of Hydrogen Sulfide and Cysteine Excreted by Bacterial Cells
    Authors:  Sergey Korshunov and James A. Imlay, date: 05/20/2018, view: 2357, Q&A: 0
    [Abstract] Bacteria release cysteine to moderate the size of their intracellular pools. They can also evolve hydrogen sulfide, either through dissimilatory reduction of oxidized forms of sulfur or through the deliberate or inadvertent degradation of intracellular cysteine. These processes can have important consequences upon microbial communities, because ...
    Isolation of Commensal Escherichia coli Strains from Feces of Healthy Laboratory Mice or Rats
    Authors:  Tingting Ju and Benjamin P. Willing, date: 03/20/2018, view: 3420, Q&A: 0
    [Abstract] The colonization abundance of commensal E. coli in the gastrointestinal tract of healthy laboratory mice and rats ranges from 104 to 106 CFU/g feces. Although very well characterized, the family that E. coli belongs to has a very homogeneous 16S rRNA gene sequence, making the identification from 16S rRNA ...
    Gene Dosage Experiments in Enterobacteriaceae Using Arabinose-regulated Promoters
    Authors:  Sanchari Bhattacharyya, Shimon Bershtein and Eugene I Shakhnovich, date: 07/20/2017, view: 3689, Q&A: 0
    [Abstract] This protocol is used to assay the effect of protein over-expression on fitness of E. coli. It is based on a plasmid expression of a protein of interest from an arabinose-regulated pBAD promoter followed by the measurement of the intracellular protein abundance by Western blot along with the measurement of growth parameters of E. coli ...
    Determination of Survival of Wildtype and Mutant Escherichia coli in Soil
    Authors:  Yinka Somorin and Conor O'Byrne, date: 07/20/2017, view: 3817, Q&A: 0
    [Abstract] E. coli resides in the gastrointestinal tract of humans and other warm-blooded animals but recent studies have shown that E. coli can persist and grow in various external environments including soil. The general stress response regulator, RpoS, helps E. coli overcome various stresses, however its role in soil survival ...
    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 ...
    Calculation of Microorganism Lag Times as a Measure of Adaptative Capability between Different Growth Conditions
    Author:  Brice Enjalbert, date: 02/05/2016, view: 3744, Q&A: 0
    [Abstract] This protocol has been designed as a simple and efficient way to investigate microorganism adaptive capabilities (Enjalbert et al., 2015). It is performed using switch experiments in which cells are initially grown in the first condition (primary cultures), then rapidly switched to the second condition (secondary culture) without ...
    Measurement of Proton-driven Antiport in Escherichia coli
    Authors:  Scarlett R. Holdsworth and Christopher J. Law, date: 11/05/2014, view: 5816, Q&A: 0
    [Abstract] Secondary active transport of substrates across the inner membrane is vital to the bacterial cell. Of the secondary active transporter families, the ubiquitous major facilitator superfamily (MFS) is the largest and most functionally diverse (Reddy et al., 2012). Recently, it was reported that the MFS multidrug efflux protein MdtM from ...



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