Secreted Expression of Mature Peptide Gene of Human Bone Morphogenic Protein-7 in Pichia pastoris

Objective: To construct and express mature peptide of human bone morphogenic protein-7(hBMP-7) in Pichia pastoris. Methods: A pair of primers based on mature peptide of hBMP-7 gene sequence were designed according to GenBank(Accession No.NM_001719). The DNA sequence encoding the mature peptide of hBMP-7 was amplified by PCR and cloned into P.pastoris expression vector pPIC9K. The recombinant pPIC9K-hBMP7 was linearized and electroporated into P.pastoris SMD1168 strain. Then expression was induced by methanol at 30℃ in secreted form. The expressing strain was selected and assayed. Results: Cloned gene was expressed in soluble form by secreting into culture medium. Recombinant protein was of 8% total secreted protein and easily identified by Western blot and ELISA with specific antibody binding activity. Conclusion: The successful cloning and expression of rhBMP-7 mature peptide in P.pastoris are conductive to further study of its function.