Site-Directed Mutagenesis and Its Application in Studying the Interactions of T3S Components
2017 (English)In: Type 3 Secretion Systems: Methods and Protocols / [ed] Matthew L. Nilles and Danielle L. Jessen Condry, Springer-Verlag New York, 2017, 11-31 p.Chapter in book (Refereed)
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.
Place, publisher, year, edition, pages
Springer-Verlag New York, 2017. 11-31 p.
, Methods in Molecular Biology, ISSN 1064-3745 ; 1531
Site-directed mutagenesis, Type III secretion systems, Suicide vector pDM4, Mutant libraries, Genetic-based screens, Protein-protein interaction assays
Biochemistry and Molecular Biology
Research subject Microbiology
IdentifiersURN: urn:nbn:se:umu:diva-128091DOI: 10.1007/978-1-4939-6649-3_2PubMedID: 27837478ISBN: 978-1-4939-6647-9ISBN: 978-1-4939-6649-3OAI: oai:DiVA.org:umu-128091DiVA: diva2:1048941
FunderSwedish Research Council, 2014–2105