Figure 4
Characterization and purification of the mutant FlhF proteins. (A) Cells of BL21(DE3) harboring a plasmid pTSK110(WT) or the derivative mutant plasmids carrying flhF-his6 in pCold IV vector were broken in the presence of GDP. The proteins were separated by SDS-PAGE and the gels were stained by CBB. The regions of interest were cropped from the gels. 1: the pre-induction cells, 2: the post-induction cells, 3: the low speed centrifugal precipitation, 4: the low speed supernatant. (B) BL21(DE3) cells harboring a pTSK110 derivative mutant plasmid (E440K) were broken in the presence of GDP. 1: the pre-induction cells, 2: the post-induction cells, 3: the low speed centrifugal precipitation, 4: the low speed supernatant, 5: the ultracentrifugation precipitation, 6: the post-ultracentrifugation supernatant, 7: the flow through fraction of Ni-NTA resin, 8: the washed fraction of Ni-NTA resin, 9: the eluted fraction of Ni-NTA resin. The proteins were separated by SDS-PAGE and the gels were stained by CBB.
