Transposon Mutagenesis in Streptococcus Species
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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.
|Title of host publication||Microbial Transposon Mutagenesis : Protocols and Applications|
|Number of pages||11|
|Publication status||Published - 2019|
|Series||Methods in Molecular Biology|
- In vivo transposon mutagenesis, Mariner, pMN100, Streptococci