[Human bone morphogenetic protein-2: recombinant expression in E. coli].
OBJECTIVE
To explore the way of producing human bone morphogenetic protein-2(hBMP-2) for bone healing by using the gene engeneering techniques.


METHODS
hBMP-2 cDNA fragment, which consists of 3' end partial propeptide and mature peptide sequence, was inserted into the multiple cloning site of expression vector pkpL-3a via ligation. The recombinant plasmid pkpL-3a/hBMP-2 was transformed into E. coli pOp2136. By the method of restriction map, the positive expression clone was selected. SDS-PAGE analysis showed a new foreign protein band near 27,000 after induction. The yield of induced hBMP-2 accouned for 10% of the total bacterial proteins.


RESULTS
The partial purified recombinant hBMP-2 was implanted into Wistar rat thigh. After 4 weeks, histological analysis showed that it induced the proliferation of mesenchymal type cells and formation of new cartilage and bone in the implantation area.


CONCLUSIONS
The hBMP-2 produced by gene engeneering techniques has the biologic capacity of ectopic bone formation.
Department of Orthopaedics, Third Teaching Hospital, Beijing Medical University.
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D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism*; D005260:Female; D006133:Growth Substances / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D006801:Humans; D008297:Male; D010012:Osteogenesis / Q000187:drug effects; D015202:Protein Engineering; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000096:biosynthesis; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D006133:Growth Substances; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2