Development of a replicating plasmid based on the native oriC in Mycoplasma pneumoniae

• DOI: 10.1099/mic.0.000711
• Publication Date: 2018-09-25T12:12:47Z
• Authors: Cedric Blötz; Carole Lartigue; Yanina Valverde T...; Estelle Ruiz; Bernhard Paetzold; Julia Busse; Jörg Stülke
• Keywords: Mycoplasma genitalium; DnaA

Bacteria of the genus Mycoplasma have recently attracted considerable interest as model organisms in synthetic and systems biology. In particular, pneumoniae is one most intensively studied field However, genetic manipulation these bacteria often difficult due to lack efficient some intrinsic peculiarities such an aberrant code. One major disadvantage working with M. replicating plasmids that can be used for complementation mutants expression proteins. this study, we analysed genomic region around gene encoding replication initiation protein, DnaA, detected putative binding sites DnaA (DnaA boxes) are, however, less conserved than other bacteria. The construction several encompassing allowed selection plasmid pGP2756 stably inherited experiments, shown by assays glpQ glycerophosphoryl diester phosphodiesterase. Plasmid-borne mutant restored formation hydrogen peroxide when were cultivated presence glycerol phosphocholine. Interestingly, also close relative, genitalium but not more distantly related members Mycoplasma. Thus, a valuable tool analysis genitalium.