Prokaryotes

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    Protocols in Current Issue
    Induction of Natural Competence in Genetically-modified Lactococcus lactis
    Authors:  Joyce Mulder, Michiel Wels, Oscar P. Kuipers, Michiel Kleerebezem and Peter A. Bron, date: 07/05/2018, view: 2528, Q&A: 0
    [Abstract] Natural competence can be activated in Lactoccocus lactis subsp lactis and cremoris upon overexpression of ComX, a master regulator of bacterial competence. Herein, we demonstrate a method to activate bacterial competence by regulating the expression of the comX gene by using a nisin-inducible promoter in an L. ...
    Targeted Genome Editing of Virulent Phages Using CRISPR-Cas9
    Authors:  Marie-Laurence Lemay, Ariane C. Renaud, Geneviève M. Rousseau and Sylvain Moineau, date: 01/05/2018, view: 7242, Q&A: 1
    [Abstract] This protocol describes a straightforward method to generate specific mutations in the genome of strictly lytic phages. Briefly, a targeting CRISPR-Cas9 system and a repair template suited for homologous recombination are provided inside a bacterial host, here the Gram-positive model Lactococcus lactis MG1363. The CRISPR-Cas9 system is ...
    Determination of Quinone Reductase Activity
    Authors:  Stefano Mancini and Marc Solioz, date: 09/05/2015, view: 5462, Q&A: 0
    [Abstract] We recently demonstrated the presence of a quinone detoxification pathway present in Firmicutes. It is based on two enzyme activities, namely a quinone reductase, YaiB, described here, and a hydroquinone dioxygenase, YaiA, described in a separate protocol. In Lactococcus lactis (L. lactis), these enzymes are encoded by the ...
    Determination of Hydroquinone Dioxygenase Activity
    Authors:  Stefano Mancini and Marc Solioz, date: 09/05/2015, view: 4868, Q&A: 0
    [Abstract] We recently demonstrated the presence of a quinone detoxification pathway present in Firmicutes. It is based on two enzyme activities, namely a hydroquinone dioxygenase, YaiA, described here, and a hydroquinone reductase, YaiB, described in a separate protocol. In Lactococcus lactis (L. lactis), these enzymes are encoded by the ...
    Amino Acid Transport Assays in Resting Cells of Lactococcus lactis
    Authors:  Hein Trip and Juke S. Lolkema, date: 06/20/2013, view: 6319, Q&A: 0
    [Abstract] Many bacteria are auxotrophic for at least a number of amino acids for which they lack the biosynthetic pathways. The organisms are still able to grow in media containing free amino acids as the sole source of amino acid when transport systems for the free amino acids are present in the cytoplasmic membrane. A range of transport systems for ...



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