Prokaryotic Expression and Purification of BIG-3 Fusion Protein and the Antibody’s Preparation and Identification

Prokaryotic expression and purification of fusion protein coded by bone morphogenetic protein induced gene(BIG-3) and preparation of its polyclonal antibody,BIG-3 gene was inserted into the expression vector pGEX-4T-2 to get recombinant plasmid pGEX-4T-2-BIG-3. The recombinant plasmid pGEX-4T-2-BIG-3 was transformed into E. coli BL 21 and fusion protein expression was induced by IPTG. New Zealand rabbits were immunized with GST-BIG-3 fusion proteins to prepare a polyclonal antibody of BIG-3. GST-BIG-3 antiserum was obtained and polyclonal antibody was characterized by western blot.It is resulted that GST-BIG-3 fusion protein was effectively expressed in E.coli. After purification, a single clear band of GST-BIG-3 fusion protein appeared in SDS-PAGE gel. Polyclonal antibody prepared from purified fusion protein has obvious characteristic.It is conclused that GST-BIG-3 fusion protein can be effectively expressed in E.coli, SDS-PAGE and Western blot analysis showed that purified GST-BIG-3 fusion protein and polyclonal antibody with characteristics was obtained.