Purification of Trx-AfusinC.

(A) SDS-PAGE analysis of the recombinant fusion protein (Trx-AfusinC) in E. coli. Expression E. coli BL21(DE3) harbouring pET-32c_afuC was induced with IPTG (+). The crude extract of induced culture is compared with non-induced culture (-). (B) The crude extract was centrifuged, and soluble (S) and insoluble (I) fractions were analysed. (C) Insoluble fraction was collected (1). Then, Inclusion bodies were washed with 1% Triton X-100 (2) and solubilised with 8 M urea (3). Solubilised Trx-AfusinC was purified by IMAC (Ni-IDA). Flow-through (4), wash (5), and elution (6) fractions are shown. Protein samples were separated on 12% Glycine SDS-PAGE gel which was stained with Coomassie blue R-250. The relative molecular weights expressed in kDalton (kDa) of the protein marker (M) are shown.