In vivo Salmonella Typhimurium Infection Protocol

In all experiments, Salmonella Typhimurium SL1344 (S.Tm;SB300; SmR) or the indicated ssaV mutant version (S.Tm^ssaV; M2730; AmpR) were used. [44 (link),45 (link)]. WITS-tags were introduced into S.Tm by P22 phage transduction and subsequent selection on kanamycin. The presence of the correct WITS-tag was confirmed by PCR using tag-specific primers [14 (link),46 (link)]. For in vivo mouse infections, bacteria were grown in lysogeny broth (LB) containing the appropriate antibiotics (50 μg/ml streptomycin (AppliChem); 15 μg/ml chloramphenicol (AppliChem); 50 μg/ml kanamycin (AppliChem); or 100 μg/ml ampicillin (AppliChem)) at 37°C for 12h and sub-cultured in 1:20 LB without antibiotics for 4h. Cells were washed and re-suspended in cold PBS (BioConcept).