Generation of Mycoplasma hominis mutants by targeting-induced local lesions in genomes (TILLING)

Mycoplasma hominis is a human urogenital pathogen for which no genetic tools are currently available. Targeting-Induced Local Lesions IN Genomes (TILLING) is a reverse-genetic method that combines point mutations induced by chemical mutagenesis with a DNA screening technique. We used ethylmethanesulfonate (EMS) to generate a library of M. hominis mutants.A 1,200-clone library was generated by submitting M. hominis PG21 to 9 mg/mL EMS for 3h.A total of four vaa- and two oppA-mutants harboring missense mutations were obtained and fully sequenced.