Prokaryotes

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    Protocols in Current Issue
    Measurement of Cellular Copper in Rhodobacter capsulatus by Atomic Absorption Spectroscopy
    Authors:  Petru-Iulian Trasnea, Dorian Marckmann, Marcel Utz and Hans-Georg Koch, date: 10/05/2016, view: 5622, Q&A: 0
    [Abstract] Copper is an essential micronutrient and functions as a cofactor in many enzymes such as heme-Cu oxygen reductases, Cu-Zn superoxide dismutases, multi-copper oxidases and tyrosinases. However, due to its chemical reactivity, free copper is highly toxic (Rae et al., 1999) and all organisms use sophisticated machineries for controlling ...
    Surface Polysaccharide Extraction and Quantification
    Authors:  Cedric Arthur Brimacombe and John Thomas Beatty, date: 10/20/2013, view: 12614, Q&A: 1
    [Abstract] Gram-negative bacterial cells possess two membranes - the inner cytoplasmic membrane and the outer membrane. The two membranes are distinct in their composition; the inner membrane is composed of a phospholipid bilayer, whereas the outer membrane (OM) is composed of an asymmetrical bilayer, with the outer leaflet containing lipopolysaccharide ...
    Bacterial Conjugation in Rhodobacter capsulatus
    Authors:  Molly M. Leung and John Thomas Beatty, date: 07/05/2013, view: 8930, Q&A: 0
    [Abstract] Bacterial conjugation of plasmids is the common method of introducing foreign DNA into Rhodobacter capsulatus because transformational systems have not been shown as efficient methods of introducing DNA to R. capsulatus. For R. capsulatus bacterial conjugation using an Escherichia coli donor can be used to ...
    Rhodobacter capsulatus Gene Transfer Agent Transduction Assay
    Authors:  Molly M. Leung and John Thomas Beatty, date: 02/20/2013, view: 7370, Q&A: 0
    [Abstract] The gene transfer agent (GTA) is a bacteriophage-like particle that transfers genomic DNA from a donor to a recipient bacterium. The Rhodobacter capsulatus GTA (RcGTA) was the first to be studied and this protocol has been optimized for RcGTA transduction, although it could be modified for other bacteria containing a GTA. The RcGTA ...
    Construction of Chromosomal In-Frame Deletion Mutants in Rhodobacter capsulatus
    Authors:  Molly M. Leung and John Thomas Beatty, date: 02/20/2013, view: 9546, Q&A: 0
    [Abstract] Markerless chromosomal in-frame deletion mutants are often used to avoid polar effects, conserve antibiotic resistance markers or make multiple mutations. This protocol capitalizes on the sacB gene (lethal in the presence of sucrose), for selection against a single cross-over event, on a suicide vector (contains no Rhodobacter capsulatus ...
    Rhodobacter capsulatus Gene Transfer Agent (RcGTA) Activity Bioassays
    Authors:  Alexander P. Hynes and Andrew S. Lang, date: 01/20/2013, view: 7559, Q&A: 0
    [Abstract] RcGTA, a small phage-like particle produced by Rhodobacter capsulatus, was initially identified in culture filtrates as a DNase-resistant form of DNA transfer between R. capsulatus cells. This gene transfer assay has been used to identify RcGTA-producing strains and to help determine the roles of genes thought to be responsible ...



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