Figure 2

Enhanced aggregation of EGFP in fusion with MoaD. In the upper panel, the recombinant proteins of EGFP fused with MoaD at N-terminus (A) or C-terminus (B), were induced by 1 mM IPTG for 4 h at 37°C. EGFP alone without fusion, and fusions with ThiS were used as control. Total cell lysate (T) and the soluble (S) or insoluble (I) fractions were resolved on 12% SDS-PAGE. All the proteins were mainly expressed in the inclusion bodies. In the lower panel, the cells were induced with 0.1 mM IPTG at room temperature for 20 h. Unboiled total cell lysates of fusions at N-terminus (C) or C-terminus (D) were resolved on 12% SDS-PAGE. The soluble native form was separated from insoluble denatured form for each sample (a). Their ratios were calculated and compared to that of EGFP (b), *p < 0.05 for triplicate experiments; # p < 0.05 for comparison between ThiS- and MoaD-fusion. The Western blot with His-tag antibody (c) and UV illuminated gel (d) further confirmed the identities of overexpressed products and corresponding native forms.