Figure 1. Production of recombinant peptide-fusion protein (PG1-MAP30-PLSN) in E. coli as inclusion bodies.

(A) Design of peptide-fusion protein: PG1 peptide was joined with N-terminal of MAP30 by 10-amino-acid linkers (underlined) and PLSN peptide was joined to the C-terminal of MAP30 by similar linkers. (B) The peptide-fusion protein was produced insolubly as inclusion bodies: Lane 1, before induction with IPTG; Lane 2, expression of peptide-fusion protein after induction; Lane 3, expression of MAP30 after induction. (C) Isolation of inclusion bodies by multiple washing steps: Lane 1, peptide-fusion protein; Lane 2, MAP30. (D) Inclusion bodies were solubilized and refolded in an alkaline buffer containing redox agents: Lane 1, peptide-fusion protein; Lane 2, MAP30.