Transposon Mutagenesis in Streptococcus Species

Martin Nilsson, Michael Givskov, Tim Tolker-Nielsen*

*Corresponding author af dette arbejde

    Abstract

    Mutant libraries, generated by transposons and screened for various phenotypes, have led to many important discoveries regarding gene functions in various organisms. In this chapter we describe the use of plasmid pMN100, a transposon vector constructed to perform in vivo transposition primarily in oral streptococci. Compared to in vitro transposition systems the conditional replicative features of the plasmid, and the inducible expression of the mariner Himar1 transposase, makes pMN100 particularly useful for bacterial strains showing a low transformation frequency. We outline how to transform plasmid pMN100 into Streptococcus mutans, carry out transposon mutagenesis, and determine the chromosomal location of inserted transposons. It is our prospect that the protocols can be used as guidelines for transposon mutagenesis in S. mutans as well as other species of streptococci.

    OriginalsprogEngelsk
    TitelMicrobial Transposon Mutagenesis : Protocols and Applications
    Antal sider11
    ForlagHumana Press
    Publikationsdato2019
    Sider39-49
    ISBN (Trykt)978-1-4939-9569-1
    ISBN (Elektronisk)978-1-4939-9570-7
    DOI
    StatusUdgivet - 2019
    NavnMethods in Molecular Biology
    Vol/bind2016
    ISSN1064-3745

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