First steps towards the genetic manipulation of Mycoplasma agalactiae and Mycoplasma bovis using the transposon Tn4001mod

Mycoplasma agalactiae and M. bovis rank amongst the most serious pathogenic mycoplasmas infecting small ruminants and cattle, respectively. Despite considerable advances made in Mycoplasma molecular genetics in the past decade, there is still a complete lack of genetic tools to assess the pathogenic mechanisms of these two species. Studies were undertaken to develop a genetic system for the analysis of potential virulence factors of these pathogens. Transposon Tn4001mod was successfully introduced into various chromosomal sites of M. agalactiae and M. bovis with an optimal frequency of 10−6 per viable colony-forming unit (CFU). This is the first report that demonstrates the amenability of these agents to transformation and to genetic manipulation. Furthermore, Tn4001 is implicated as the first potential genetic tool available for these ruminant pathogens.