Purification and Characterization of Glutathione Binding Protein GsiB from Escherichia coli

Figure 1

Expression and purification of GsiB from E.coli. (a) Recombinant plasmid digestion with restriction enzymes. M: Marker; Lane 1: plasmid digested with NcoI and HindIII. (b) SDS-PAGE analysis of GsiB expression. M: Marker; Lanes 1-2: total protein and soluble fraction of GsiB induced with 0.1 mM IPTG at 22°C for 20 h. (c) Purity analysis of GsiB. The protein was separated on 12 % (v/v) SDS-PAGE and analyzed with QuantiyOne software. M: Marker; Lane 1: purified GsiB protein; (d) Western blot analysis. Lanes 1-2: total protein and soluble fraction of GsiB in BL21 (DE3) grown at 22°C for 20 h induced with 0.1 mM IPTG; Lane 3: total protein of GsiB in BL21 (DE3) grown at 22°C for 20 h without IPTG.

(a)

(b)

(c)

(d)