Figure 6.

In vivo glyS T-box-mediated transcription antitermination assay. (A) Schematic representation of the glyS T-box-dependent lacZ expression in E. coli. The full length (FL) glyS T-box and the tRNA^Gly_GCC were cloned into pRB382 and pBAD18 plasmid, respectively. Both constructs or pRB382-T-box alone were transformed into a ΔlacZ E. coli strain. T corresponds to T-box terminator conformation and anti-T to T-box antiterminator conformation. Diagram of the β-galactosidase activity in the presence of tigecycline (B) and linezolid (C). The bars represent β-gal activity relative to cell density, in Miller Units. Error bars represent ± SD values from three independent experiments. The β-galactosidase activity was measured after 4 h of culture in either rich (+Gly) or minimal media (–Gly) in the presence or in the absence of tigecycline or linezolid (concentrations tested half-IC₅₀, IC₅₀ and IC₉₀) and after expression induction of S. aureus tRNA^Gly_GCC.