Precise excision and secondary transposition of TnphoA in non-motile mutants of a Salmonella enterica serovar Enteritidis clinical isolate.
Mutagenesis with TnphoA has been widely used in many bacteria. Here, we report the excision and secondary transposition of this transposon in three non-motile (fliC, fliF and motB) mutants of Salmonella enterica serovar Enteritidis (S. Enteritidis). Isolation of motile revertants showed that they were kanamycin resistant and conserved a copy of TnphoA in their genome in an insertion site different from the initial one. They also expressed an intact flagella. Characterization of the motile revertant derived from the fliC mutant showed that TnphoA excised precisely from the fliC gene, resulting in an equivalent amount of FliC secreted protein in the revertant compared to that of the wild-type strain. These results show that TnphoA mutants should be used with care and underline the value of using transposon derivatives lacking the transposase gene.
Amy
Mai Té
MT
Institut National de la Recherche Agronomique, Centre de Tours-Nouzilly, Pathologie Infectieuse et Immunologie, 37380 Nouzilly, France.
Virlogeux-Payant
Isabelle
I
Bottreau
Elisabeth
E
Mompart
Florence
F
Pardon
Pierre
P
Velge
Philippe
P
D000469:Alkaline Phosphatase; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements; D005407:Flagella / Q000502:physiology / Q000648:ultrastructure; D005408:Flagellin / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology; D009068:Movement; D011995:Recombination, Genetic; D012477:Salmonella enteritidis / Q000235:genetics* / Q000502:physiology
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C075661:Flif protein, Bacteria; D008565:Membrane Proteins; C064779:MotB protein, Bacteria; D005408:Flagellin; D018844:Cyclin-Dependent Kinases; D000469:Alkaline Phosphatase