Fig. 5.

Overexpression of the ribosome recycling factor (Ms-RRF) partially restores macrolide sensitivity of ΔMs_hflX. Complementing strains were created by integrating either Ms_RRF or Ms_hflX at the Bxb1 attB site of ΔMs_hflX. Expression of Ms-RRF in the complementing strain was verified using real-time PCR (SI Appendix, Table S2). Tenfold serial dilutions of wild-type M. smegmatis, ΔMs_hflX, and the complementing strains were grown to A₆₀₀ of 0.7 and spotted on Middlebrook 7H10 ADC containing indicated concentrations of macrolides.