| Abstract [eng] |
Genetic studies of mycoplasmas are limited by the lack of a replicable vector system. Mycoplasma gallinarum, an opportunistic pathogen having a wide range of hosts, is not the exception. This study describes the identification and cloning of the M. gallinarum origin of replication (oriC) in order to construct the first vectors for this species. Additionally, this report provides the first evidence of the successful transformation of M. gallinarum. Five out of six developed vectors have been functional. The replication of the sixth vector has not been supported due to the mutation in the AT rich region downstream the second DnaA box. During in vitro passages, some vectors have been integrated into the chromosome. The rate of chromosomal integration has been even further increased by the lack of antimicrobial selection pressure during cultivation. Hence developed oriC plasmids are useful for genetic studies, including inactivation or expression of selected genes, in M. gallinarum, and lead to a better understanding of its molecular biology. |
