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    Site-directed mutagenesis and its application in studying the interactions of T3S components

    Francis, M.S. and Amer, A.A.A. and Milton, D.L. and Costa, Tiago (2016) Site-directed mutagenesis and its application in studying the interactions of T3S components. In: Niles, M.L. and Condry, D. (eds.) Type 3 Secretion Systems: Methods and Protocols. Methods in Molecular Biology 1531. New York, U.S.: Humana Press, pp. 11-31. ISBN 9781493966479.

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    Type III secretion systems are a prolific virulence determinant among Gram-negative bacteria. They are used to paralyze the host cell, which enables bacterial pathogens to establish often fatal infections—unless an effective therapeutic intervention is available. However, as a result of a catastrophic rise in infectious bacteria resistant to conventional antibiotics, these bacteria are again a leading cause of worldwide mortality. Hence, this report describes a pDM4-based site-directed mutagenesis strategy that is assisting in our foremost objective to better understand the fundamental workings of the T3SS, using Yersinia as a model pathogenic bacterium. Examples are given that clearly document how pDM4-mediated site-directed mutagenesis has been used to establish clean point mutations and in-frame deletion mutations that have been instrumental in identifying and understanding the molecular interactions between components of the Yersinia type III secretion system.


    Item Type: Book Section
    Keyword(s) / Subject(s): Site-directed mutagenesis, Type III secretion systems, Suicide vector pDM4, Mutant libraries, Genetic-based screens, Protein-protein interaction assays
    School: Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences
    Depositing User: Administrator
    Date Deposited: 08 Mar 2017 14:08
    Last Modified: 02 Aug 2023 17:30


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