Fig. 1.
Cloning and purification of S. oneidensis RNAP. (A) Schematic diagram of the S. oneidensis RNAP co-overexpression construct. (B) SDS–PAGE gel of the purified S. oneidensis RNAP core. (Lane M) Protein molecular weight marker (New England Biolabs); (lanes 1–8) S. oneidensis RNAP core fractions from anion exchange chromatography; (lane 9) purified S. oneidensis σ70. (C) SDS–PAGE gel of IMAC-based pull-down assay from cell extract containing overexpressed α subunits. (Lane M) Protein molecular weight marker; (lane 1) S. oneidensis α overexpression; (lane 2) E. coli α overexpression; (lane 3) S. oneidensis RNAP core. (D) SDS–PAGE gel of purified σ factors. (Lane M) Protein molecular weight marker; (lane 1) σ70; (lane 2) σ38; (lane 3) σ32; (lane 4) σ24. Gels were Coomassie stained.