Cloning,expression and determination of bone inductive activity of hOP-1 mature peptide in escherichia coli

Objective:To amplify and clone the cDNA gene of human osteogenic protein 1 (hOP 1) mature peptide,then express its recombinant protein in E coli Method:The cDNA gene of hOP 1 mature peptide was amplified by polymerase chain reaction(PCR) and inserted into expression vector pDH in which foreign gene was controlled by P RP L promoters The recombinant plasmic pDOPm was transformed into E coli DH5α,and the expressed product was partially purified and then refolded,its bone inductive activity was assayed in mouse heterotopicaly in vivo Result:The cDNA gene of hOP 1 mature peptide was amplified;a new anticipated 16 ku protein band appeared on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) gel and amounted to 21% of total bacterial protein The expressed product existed in a form of inclusion body After being partially purified and refolded,hOP 1 mature peptide could heterotopically induce the formation of cartilage and bone tissue Conclusion:The cDNA gene of hOP 1 has been successfully cloned and efficiently expressed in E coli and has bone inductive activity after being refolded