Protocols in Current Issue
    A High-throughput Interbacterial Competition Platform
    Authors:  Hsiao-Han Lin and Erh-Min Lai, date: 09/05/2020, view: 616, 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: 1192, 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: 3980, 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: 4959, 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: 4398, 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 ...
    A Flow-assay for Farnesol Removal from Adherent Candida albicans Cultures
    Authors:  Melanie Polke and Ilse D. Jacobsen, date: 10/05/2017, view: 4824, Q&A: 0
    [Abstract] Here, we describe a protocol for a continuous flow system for C. albicans cultures growing adherent to a plastic surface. The protocol was adapted from a previous method established to simulate blood flow on endothelial cells (Wilson and Hube, 2010). The adapted protocol was used by us for the removal of molecules in C. albicans ...
    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 ...
    Protein Expression Protocol for an Adenylate Cyclase Anchored by a Vibrio Quorum Sensing Receptor
    Authors:  Stephanie Beltz and Joachim E. Schultz, date: 01/20/2017, view: 5922, Q&A: 0
    [Abstract] The direct regulation of a mycobacterial adenylate cyclase (Rv1625c) via exchange of its membrane anchor by the quorum sensing receptor CqsS (Vibrio harveyi) has recently been reported (Beltz et al., 2016). This protocol describes the expression and membrane preparation for these chimeric proteins.
    In vitro Autophosphorylation and Phosphotransfer Assay of Cyanobacterial Histidine Kinase 2
    Author:  Iskander M. Ibrahim, date: 12/05/2016, view: 7678, Q&A: 0
    [Abstract] This is a detailed protocol of an autophosphorylation and phosphotransfer activities of Synechocystis sp. PCC 6803 full-length Histidine Kinase 2 (Hik2) protein described by Ibrahim et al., 2016. In this protocol, radioactively labelled ATP was used to study an autophosphorylation and phosphotransfer activity of the full-length ...
    Separation of Free and Bound cAMP in Mycobacteria
    Authors:  Arka Banerjee and Sandhya S. Visweswariah, date: 07/20/2016, view: 5635, Q&A: 0
    [Abstract] Mycobacterial genomes encode a plethora of genes that are involved in the synthesis, utilization and degradation of cAMP. The genome of M. tuberculosis H37Rv, for example, encodes 16 adenylyl cyclases and 10 genes harbouring the cyclic nucleotide-binding (CNB) domain (Shenoy and Visweswariah, 2006). Cyclic AMP is efficiently secreted by ...

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