title: The presence of the tet gene from cloning vectors impairs Salmonella survival in macrophages.
abstract: Cloning, mutagenesis and complementation of virulence factors are key steps to understand the mechanisms of bacterial pathogenesis and cloning vectors are routinely utilized for these processes. We have investigated the effect of the presence of commonly used cloning vectors on the survival of the intracellular bacterial pathogen Salmonella during macrophage infection. We demonstrate that the presence of the pSC101 derived tetracycline resistance gene on plasmids causes a lower survival rate of Salmonella in macrophages. The decrease in survival caused by the presence of the tet gene was not due to a higher susceptibility to gentamicin, a growth defect, or to increased sensitivity to acid. Higher susceptibility to hydrogen peroxide was observed in vitro for strain containing plasmid with the tet gene when the strains were grown at high densities but not when they were grown at low densities. Our findings demonstrate that the use of the tet gene for mutation or complementation can have deleterious effects and should thus be carefully considered.
authors.affiliation: Washington University, USA.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Abromaitis
authors.forename: Sébastien
authors.initials: S
authors.lastname: Faucher
authors.forename: Maxime
authors.initials: M
authors.lastname: Béland
authors.forename: Roy
authors.initials: R
authors.lastname: Curtiss
authors.forename: France
authors.initials: F
authors.lastname: Daigle
mesh_terms: D018929:Cell Culture Techniques; D003001:Cloning, Molecular; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D008264:Macrophages / Q000382:microbiology* / Q000473:pathology; D010957:Plasmids / Q000235:genetics*; D012475:Salmonella / Q000235:genetics* / Q000502:physiology; D013753:Tetracycline Resistance / Q000235:genetics*