FIG. 1.
SDS-PAGE analysis of purified TxAC314HA. E. coli BL21(DE3) carrying the plasmid pGEX/TxAC314 was grown for 6 h at 37°C following induction with IPTG; cells were then collected by centrifugation and lysed. Soluble proteins were subjected to affinity chromatography purification on a glutathione-Sepharose 4B resin and then separated through SDS-12% PAGE gels stained with Coomassie blue. Lane A, total cell lysate following 6 h of induction with IPTG and before affinity column purification; lane B, recombinant protein (GST-TxAC314) obtained following affinity chromatography of cell lysate on glutathione-Sepharose 4B resin and collected by addition of reduced glutathione; lane C, purified GST and TxAC314 obtained following digestion of recombinant GST-TxAC314 with thrombin; lane D, purified recombinant TxAC314 obtained by overnight digestion with thrombin (1 U/20-ml resin bed volume) of cell lysate purified and immobilized on glutathione-Sepharose 4B resin; lane MW, molecular mass markers (apparent molecular masses are shown in daltons).