Ribosome rescue through the trans-translation pathway, which ensure translational fidelity, is a novel antibacterial drug target. Ribosomal protein L9 enhances the fidelity of protein translation in bacteria, which may affect ribosome rescue from trans-translation. The impact of ribosomal protein L9 on the activity of known inhibitors of trans-translation was investigated using wild-type and L9-deletion strains of E. coli. Growth inhibitory analysis from the two strains suggest that ribosomal protein L9 functions independent of trans-translation.