Microbiology

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    Protocols in Current Issue
    Visualization of Host Cell Kinase Activation by Viral Proteins Using GFP Fluorescence Complementation and Immunofluorescence Microscopy
    Authors:  Sherry T. Shu, Wing Fai Li and Thomas E. Smithgall, date: 07/05/2021, view: 1418, Q&A: 0
    [Abstract]

    Non-receptor protein-tyrosine kinases regulate cellular responses to many external signals and are important drug discovery targets for cancer and infectious diseases. While many assays exist for the assessment of kinase activity in vitro, methods that report changes in tyrosine kinase activity in single cells have the potential to provide

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    Analysis of Pseudomonas aeruginosa c-di-GMP High and Low Subpopulations Using Flow-assisted Cell Sorting (FACS) and Quantitative Reverse Transcriptase PCR (qRT-PCR)
    Authors:  Catherine R. Armbruster and Matthew R. Parsek, date: 01/20/2021, view: 1115, Q&A: 0
    [Abstract]

    Cyclic diguanylate monophosphate (c-di-GMP) is a second messenger signaling molecule that drives the transition from planktonic to the biofilm mode of growth in many bacterial species. Pseudomonas aeruginosa has at least two surface sensing systems that produce c-di-GMP in response to surface attachment, the Wsp and Pil-Chp systems. We recently

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    Assessment of Diadenylate Cyclase and c-di-AMP-phosphodiesterase Activities Using Thin-layer and Ion Exchange Chromatography
    Authors:  Andreas Latoscha, David Jan Drexler, Gregor Witte and Natalia Tschowri, date: 01/05/2021, view: 1019, Q&A: 0
    [Abstract]

    All living cells use cyclic nucleotides as second messengers for signal sensing and transduction. Cyclic di-3′,5′-adenosine monophosphate (c-di-AMP) is primarily involved in the control of bacterial and euryarcheal osmoadaptation and is produced by diadenylate cyclases from two molecules of ATP. Specific phosphodiesterases hydrolyze

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    Candida albicans Culture, Cell Harvesting, and Total RNA Extraction
    Authors:  Max V. Cravener and Aaron P. Mitchell, date: 11/05/2020, view: 1344, Q&A: 0
    [Abstract]

    Transcriptional analysis has become a cornerstone of biological research, and with the advent of cheaper and more efficient sequencing technology over the last decade, there exists a need for high-yield and efficient RNA extraction techniques. Fungi such as the human pathogen Candida albicans present a unique obstacle to RNA purification in the

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    Measurement of Ascorbic Acid and Glutathione Content in Cyanobacterium Synechocystis sp. PCC 6803
    [Abstract] Ascorbic acid (AsA) and gluthathione (GSH) are two key components of the antioxidant machinery of eukaryotic and prokaryotic cells. The cyanobacterium Synechocystis sp. PCC 6803 presents both compounds in different concentrations (AsA, 20-100 μM and GSH, 2-5 mM). Therefore, it is important to have precise and sensitive methods to ...
    A High-throughput Interbacterial Competition Platform
    Authors:  Hsiao-Han Lin and Erh-Min Lai, date: 09/05/2020, view: 1946, Q&A: 0
    [Abstract] Contact-dependent interbacterial competition is a common strategy used by bacteria to fight for their ecological niches. Interbacterial competition is monitored by a competition assay involving co-culturing the attacker and the recipient bacterial cells on agar, followed by recovery of the surviving recipient cells. Conventional interbacterial ...
    Growth Recovery Assay and FACS-based Population Sorting Following Territorial Exclusion in Proteus mirabilis
    Authors:  Murray J. Tipping and Karine A. Gibbs, date: 03/05/2020, view: 2249, Q&A: 0
    [Abstract] Many bacteria take part in self recognition and kin discrimination behavior using contact-dependent effectors. Understanding the effects these effectors cause is important to explain bacterial community formation and population dynamics. Typically, kin discrimination effectors are toxins that kill target cells; their effect is therefore obvious ...
    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: 5068, 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 ...
    Bacterial Competition Assay Based on Extracellular D-amino Acid Production
    Authors:  Laura Alvarez and Felipe Cava, date: 04/05/2018, view: 6023, 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 ...
    In vivo Analysis of Cyclic di-GMP Cyclase and Phosphodiesterase Activity in Escherichia coli Using a Vc2 Riboswitch-based Assay
    Authors:  Ying Liu, Hyunhee Kim and Ute Römling, date: 03/05/2018, view: 5321, Q&A: 0
    [Abstract] Cyclic di-guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger that regulates distinct aspects of bacterial physiology. It is synthesized by diguanylate cyclases (DGCs) and hydrolyzed by phosphodiesterases (PDEs). To date, the activities of DGC and PDE are commonly assessed by phenotypic assays, mass spectrometry analysis of ...



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