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GacI and GacJ exist as a detergent-stable complex in the membrane. GacJ and His-tagged GacI were co-expressed in E. coli Rosetta DE3 cells and extracted from the membrane fraction in 2.5% CHAPS. The proteins were purified using Ni-NTA-agarose in the presence of 2.5% CHAPS. A, fractions collected during Ni-NTA purification were analyzed by immunoblot using anti-His antibodies. B, eluted proteins were analyzed by SDS-PAGE. The results are representative of three separate experiments.
