Transposon mutagenesis in Mycoplasma hyopneumoniae using a novel mariner-based system for generating random mutations

摘要: Mycoplasma hyopneumoniae is the cause of enzootic pneumonia in pigs, a chronic respiratory disease associated with significant economic losses to swine producers worldwide. The molecular pathogenesis infection poorly understood due lack genetic tools allow manipulation organism and more generally for genus. objective this study was develop system generating random transposon insertion mutants M. that could prove powerful tool enabling be unraveled. A novel delivery vector constructed containing hyperactive C9 mutant Himar1 transposase along mini tetracycline resistance cassette, tetM. strain 232 electroporated construct tetM-expressing transformants selected on agar tetracycline. Individual contained single insertions were stable upon serial passages broth medium. sites 44 individual determined confirmed disruption several genes. large pool over 10 000 generated should saturation genome. This first time mutagenesis has been demonstrated important pathogen applied other species are intractable manipulation. ability generate libraries further pathogen.