Random transposon insertion in the Mycoplasma hominis minimal genome

摘要: Mycoplasma hominis is an opportunistic human pathogen associated with genital and neonatal infections. Until this study, the lack of a reliable transformation method for genetic manipulation M. hindered investigation pathogenicity peculiar arginine-based metabolism bacterium. A genomic analysis 20 different strains revealed number putative restriction-modification systems in species. Despite presence these systems, reproducible polyethylene glycol (PEG)-mediated protocol was successfully developed study three strains: two clinical isolates M132 reference strain. Transformants were generated by transposon mutagenesis efficiency approximately 10-9 transformants/cell/µg plasmid shown to carry single or multiple mini-transposons randomly inserted within their genomes. One M132-mutant observed single-copy gene encoding P75, protein potentially involved adhesion. However, no difference adhesion cell-assays between mutant parent Whole genome sequencing mutants carrying copies further occurrence rearrangements. Overall, first time that genetically modified have been obtained random using mini-transposon conferring resistance tetracycline.