Figure S1. Clustal W multiple alignment of SigA, SigB, and SigF sigma factor proteins.

1. R2 R4 Figure S1. Clustal W multiple alignment of SigA, SigB, and SigF sigma factor proteins. Figure S1. Clustal W multiple sequence alignment of SigA, SigB, and SigF sigma factor proteins. The amino acid sequences of SigA, SigB, and SigF were obtained from TubercuList database (http://genolist.pasteur.fr/TubercuList/) and aligned with Clustal W multi-alignment sequence analysis program. The boxes indicate R2 and R4 regions, which are known promoter binding regions at -10 and -35, respectively.

2. Figure S2.    Growth rates of sigB and sigF knock-in strains in vitro. Figure S2. Growth rates of the sigB and sigF knock in strains in vitro. The effect of sigma factor overexpression on the in vitro growth of M. tuberculosis in complete Middlebrook 7H9 medium was investigated by growing M. tuberculosis harboring the empty vector pSCW38 (P), the sigB knock-in vector (pSCW40, pB), and the sigF knock-in vector (pSCW35, pF) to an A600 of 0.5 and then inducing knock-in expression by the addition of acetamide to 0.2%. Growth rate was followed by monitoring A600 over time. ○:Control, △: sigB KI, □ sigF KI.

3. Figure S3. SDS-PAGE of purified SigB and SigF proteins. kDa 2 1 50 20 Figure S3. SDS-PAGE of purified SigB and SigF proteins. His-tagged versions of the SigB and SigF proteins were overexpressed in E. coli with T7 RNA polymerase overexpression system and purified with nickel affinity chromatography. The purified proteins were identified by SDS-PAGE with commercially-available size markers (Invitrogen). 1. Purified SigB, 2. Purified SigF