Prokaryotic expression, protein purification of a protein phosphatase 2C gene (ZmPP2C2) from maize

The coding region of the protein phosphatase 2C gene (ZmPP2C2) from Zea mays (maize) was sub-cloned into expression vector, pET30a-c(+), and introduced into E. coli BL21 (DE3) for expression. SDS-PAGE analysis indicated ZmPP2C2 was great expressed at 37°C for 4 h with 1mM IPTG. Ultrasonic extraction experiment showed this recombinant protein was little in lysis buffer solution, but most in inclusion body. Inclusion body was collected and dissolved by lysis buffer solution with carbamide. The hexahistidine-tagged ZmPP2C2 fusion protein was purified by Ni-NTA column by 300 mM imidazole elution.