Expression and purification of TfRscFv-GAL4 fusion protein. A, SDS-PAGE showing the fusion protein levels present in the inclusion bodies after IPTG induction at different temperature. BL21 E.coli cell were transformed with TfRscFv-GAL4-pET plasmid and gene expression was showed that Inclusion bodies extract after IPTG induced 24 hours at 37°C (Lane1), 30°C (Lane2), 23°C (Lane3). Lane M: protein molecular weight marker. B, after IPTG induced 24 hours at 30°C at different time, Western blotting analysis TfRscFv-GAL4 fusion protein purified from inclusion bodies by IMAC, as showed in 0h (lane 1), 4h (lane 2), 6h (lane 3), and BL21E.coli transformed with an empty vector (lane 4). The highest level of protein expression was at 6h after induction.