Figure 2
Purification of FlhF. (A) Cells of BL21(DE3) harboring a plasmid pTSK110 which has flhF-his6 in pCold IV vector were broken in the presence of GDP, GTP, ADP and ATP with (lower panel) or without (upper panel) MgCl2. 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) Purification of FlhF by adding GTP or 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 eluted fraction of Ni-NTA resin. The proteins were separated by SDS-PAGE and the gels were stained by Coomassie brilliant blue (CBB).
