Genome Engineering in Mycoplasma gallisepticum Using Exogenous Recombination Systems
journal contributionposted on 2022-02-15, 23:02 authored by Thomas Ipoutcha, Géraldine Gourgues, Carole Lartigue, Alain Blanchard, Pascal Sirand-Pugnet
Mycoplasma gallisepticum (Mgal) is a common pathogen of poultry worldwide that has recently spread to North American house finches after a single host shift in 1994. The molecular determinants of Mgal virulence and host specificity are still largely unknown, mostly due to the absence of efficient methods for functional genomics. After evaluating two exogenous recombination systems derived from phages found in the phylogenetically related Spiroplasma phoeniceum and the more distant Bacillus subtilis, the RecET-like system from B. subtilis was successfully used for gene inactivation and targeted replacement in Mgal. In a second step, the Cre-lox recombination system was used for the removal of the antibiotic resistance marker in recombinant mutants. This study therefore describes the first genetic tool for targeted genome engineering of Mgal and demonstrates the efficiency of heterologous recombination systems in minimal bacteria.
study therefore describesstill largely unknownheterologous recombination systemsfirst genetic toolantibiotic resistance markerspiroplasma phoeniceum mycoplasma gallisepticum single host shiftlox recombination systembacillus subtilis targeted genome engineeringsubtilis genome engineeringmgal targeted replacementlike systemhost specificitysecond steprecombinant mutantsrecently spreadpoultry worldwidephylogenetically relatedphages foundmostly duemolecular determinantsminimal bacteriagene inactivationfunctional genomicsefficient methodscommon pathogen