Figure 2
Expression of recombinant protein in ZYP-5052 medium. A). Coomassie stained 12% SDS-PAGE of lysates shows proteins expressed in ZYP-5052 at 25°C at different incubation time. Equal culture densities (corresponding to 1 OD600 cells) were analysed in each lane. Cells grown to saturation in PG, a non-inducing growth medium were loaded as control. Lane L: Protein marker; Lane C: Control; Lane 3 h-45 h: Induced samples at different incubation time, as indicated on top of each lane. B). Western blot of expressed protein. Expressed protein was transferred to nitrocellulose membrane at 30 V overnight at 4°C. After blocking in 3% BSA, membrane was treated with mouse raised anti-His antibody (1 : 500) and HRP-conjugated anti-mouse IgG antibody and developed with DAB. Bold arrow denotes distinct band in induced sample corresponding to expressed protein of 25 kDa size. Control used is culture grown to saturation in PG medium. No expression is seen in control. Lane L: Protein marker; Lane C: Control; Lane 45 h: Sample processed 45 hr post incubation at 25°C.