title: Expression and purification of recombinant human bone morphogenetic protein-7 in Escherichia coli.
abstract: Bone morphogenetic protein-7 (BMP-7) is a multifunctional cytokine of the transforming growth factor β superfamily, which induces bone formation and plays an important role during bone tissue repair and embryonic development. In this study, human BMP-7 (hBMP-7) cDNA was cloned and expressed in Escherichia coli, and its yield was approximately 30% of the total bacterial protein. After the bacteria were lysed by ultrasonication and repeated washing, inclusion bodies were extracted and dissolved using a high-strength denaturant. The monomer of rhBMP-7 was purified by ion-exchange chromatography, and the purity coefficient was approximately 96%. The protein was renatured with refolding buffers at different pH values. The renatured rhBMP-7 dimer protein in this study increased the alkaline phosphatase activity of NIH3T3 cells. This study may be helpful for the in vitro production and biomedical application of rhBMP-7 protein expressed in an E. coli expression system.
authors.affiliation: a Department of Pharmacy , Wannan Medical College , Wuhu , P. R. China.
authors.forename: Guodong
authors.initials: G
authors.lastname: Wang
authors.forename: Jinxiang
authors.initials: J
authors.lastname: Han
authors.forename: Shili
authors.initials: S
authors.lastname: Wang
authors.forename: Ping
authors.initials: P
authors.lastname: Li
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D055419:Bone Morphogenetic Protein 7 / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D003001:Cloning, Molecular; D004926:Escherichia coli; D015870:Gene Expression; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: C525819:BMP7 protein, human; D055419:Bone Morphogenetic Protein 7; D011994:Recombinant Proteins; D000469:Alkaline Phosphatase