Fig. 3

Recombinant protein expression of 6xHis-Hill_BB_C10074. A SDS-PAGE of whole cell lysate from E. coli BL21 (DE3) cells transformed with T7 inducible plasmid encoding the 6xHis-Hill_BB_C10074 constructs induced (I) with 1.0 mM IPTG and uninduced (UI). B SDS-PAGE of total protein from E. coli BL21(DE3) cells induced with either 1.0 mM (A), 0.5 mM (B) or 0.1 mM (C) IPTG for 4 h. (C) Western blot using anti-6xHis antibody to detect 6xHis-Hill_BB_C10074 following expression in E. coli BL21(DE3) cells induced with either 1.0 mM (A), 0.5 mM (B) or 0.1 mM (C) IPTG for 4-hours. D SDS-PAGE of solubility fractionated total-cell protein contents following expression induced with either 1.0 mM (A), 0.5 mM (B) or 0.1 mM (C) IPTG for 4-hours to determine whether 6xHis-Hill_BB_C10074 was soluble or insoluble. E Phase-contrast visualisation of E. coli BL21(DE3) cells expressing 6xHis-Hill_BB_C10074 into inclusion bodies over the course of 4 h of induction in response to varying concentrations of IPTG (0.1 mM, 0.5 mM and 1.0 mM). Scale bar = 5 µM. F SDS-PAGE and western blot of purified 6xHis-Hill_BB_C10074. A higher MW band (indicated with an asterisk) may be a multimer of 6xHis-Hill_BB_C10074. In all images, recombinant 6xHis-Hill_BB_C10074 protein is indicated by a red arrow. Full length gels and blots are shown in Supplementary Fig. 1