FIG 1.

Roles of ω in M. tuberculosis and E. coli RNAP core assembly. (A) SDS-PAGE analysis of purified M. tuberculosis RNAP core, coreΔω, and coreΔω coexpressed with ω^Flag protein. Proteins loaded onto 4 to 20% gel to separate β and β′ subunits are shown in the middle panel. Western blots of ω^Flag with a Flag tag antibody are shown in the bottom panel. (B) Purified E. coli core and coreΔω. (C) In vitro reconstitution of M. tuberculosis and E. coli core with or without ω^His. Western blots of ω^His with a His tag antibody are shown in the bottom panel. (D) In vitro abortive transcription of purified M. tuberculosis core or coreΔω at the Rv1494 promoter (Rv1494p). RbpA and σ^A were added as indicated to reconstitute RNAP for promoter-dependent transcription. (E) In vitro abortive transcriptional activities of in vivo or in vitro reconstituted M. tuberculosis core in the presence or absence of the ω subunit at Rv1494p. RbpA and σ^A were added in each reaction in promoter-dependent transcription assay. A representative result from three independent tests is shown. Mtb, M. tuberculosis; Eco, E. coli.