Secreted expression of mature peptide of human bone morphogenetic protein-2 in Pichia pastoris

Objective To construct and express mature peptide of human bone morphogenetic protein-2 (hBMP-2) in Pichia pastoris. Methods The gene encoding of hBMP-2 was amplification by PCR and cloned into Pichia pastoris expression vector pPICZaC. The recombinant pPICZaC/hBMP2 was transformed into the Pichia pastoris X-33 strain via electroporation. Then the rhBMP-2 was expressed induced by methanol at 28℃. The high level expression was selected and assayed by the methods of PCR, SDS-PAGE and Western Blot. The rhBMP-2 was purified and the bioactivity of it was initially assayed. Results The rhBMP-2 was secreted into the supernatant and the concentration reached 100mg.L-1. The molecular mass was initially identified by SDS-PAGE. And the rhBMP-2 was further identified by Western Blot and ELISA with specific antibody binding activity. Conclusions The rhBMP-2 was successfully constructed and expressed in Pichia Pastoris. And this contributes to further study of its function and activity.