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. Author manuscript; available in PMC: 2017 Feb 1.
Published in final edited form as: Genes Cells. 2016 Jan 13;21(2):163–184. doi: 10.1111/gtc.12334

Figure 5. Complex formation requires the SSB C-terminus.

Figure 5

1L cultures of cells expressing PriA and SSBΔC8, and separately, RecG and SSBΔC8 were grown to early log phase, IPTG added to 200 µM and growth continued until early stationary phase. Cells were harvested by centrifugation, lysed and the cleared cell lysate applied to a 5ml nickel column as described in the Experimental Procedures. Proteins were then eluted using an imidazole gradient following extensive washing to remove unbound proteins. Aliquots from various fractions throughout the purification were subjected to electrophoresis. The resulting 12% SDS-PAGE gels are shown. Panels (A) and (B), helicases are N-terminal histidine tagged. Panels (C) and (D) SSBΔC8 is N-terminal histidine tagged. M, molecular weight marker; CCL, cleared cell lysate; FT, flow through. Numbers at the top of lanes indicate fraction numbers from the peak.