TY - JOUR AB - Mycoplasma hyopneumoniae is a prevalent swine respiratory pathogen that is a major cause of economic loss topig producers. Control is achieved by a combination of antimicrobials, vaccination and management practices, butcurrent vaccines offer only partial control and there is a need for improved preventative strategies. A major barrierto advances in understanding the pathogenesis of M. hyopneumoniae and in developing new vaccines is the lackof tools to genetically manipulate the organism. We describe the development and optimisation of the firstsuccessful plasmid-based system for the genetic manipulation of M. hyopneumoniae. Our artificial plasmids containthe origin of replication (oriC) of M. hyopneumoniae along with tetM, conferring resistance to tetracycline. Withthese plasmids, we have successfully transformed M. hyopneumoniae strain 232 by electroporation, generatingtetracycline resistant organisms. The persistence of extrachromosomal plasmid and maintenance of plasmid DNAover serial passages shows that these artificial plasmids are capable of self-replication in M. hyopneumoniae. Inaddition to demonstrating the amenability of M. hyopneumoniae to genetic manipulation and in optimising theconditions necessary for successful transformation, we have used this system to determine the minimum functionaloriC of M. hyopneumoniae. In doing so, we have developed a plasmid with a small oriC that is stably maintainedover multiple passages that may be useful in generating targeted gene disruptions. In conclusion, we havegenerated a set of plasmids that will be valuable in studies of M. hyopneumoniae pathogenesis and provide a majorstep forward in the study of this important swine pathogen. AU - Maglennon,GA AU - Cook,BS AU - Matthews,D AU - Deeney,AS AU - Bossé,JT AU - Langford,PR AU - Maskell,DJ AU - Tucker,AW AU - Wren,BW AU - Rycroft,AN DO - 10.1186/1297-9716-44-63 PY - 2013/// SN - 1297-9716 TI - Development of a self-replicating plasmid system for Mycoplasma hyopneumoniae T2 - Veterinary Research UR - http://dx.doi.org/10.1186/1297-9716-44-63 UR - http://hdl.handle.net/10044/1/28166 VL - 44 ER -