Production of bone morphogenetic protein-7 using pET expression system

• Published in: Current applied physics pp. 422 - 425
• Main Authors: Dong Hee Lee, Hyun Sook Baek, 이미희, 박종철
• Format: Journal Article
• Language: Korean
• Published: 한국물리학회 01.07.2005
• Subjects: 물리학
• ISSN: 1567-1739; 1878-1675

The polypeptide representing the mature part of human bone morphogenetic protein-7 (BMP-7) was eciently expressed inE. coliBL21 (DE3). The rhBMP-7 is a disulde-bonded homodimeric protein with an apparent MW of 29,000 as shown by theSDS-PAGE and gel chromatography. rhBMP-7 stimulates ALP specic activity in a dose-dependent manner. In a kinetic experi-ment, ALP activity in both the rhBMP-7 (100 ng/ml)-treated and control cultures increases gradually, and activity in therhBMP-7-treated culture is consistently higher throughout the culture period. We also quantitated the specic osteoblastic markerosteocalcin in rhBMP-7-treated culture medium. When serum-starved MC3T3-E1 cells are treated with medium containing variousconcentrations of rhBMP-7, the production of osteocalcin is increased about 7.2-fold in a dose-dependent manner up to 320 ngrhBMP-7/ml. In summary, we established anE. coliexpression system for a high level of rhBMP-7 production and inductive eectsof osteogenic dierentiation markers by rhBMP-7 in MC3T3-E1 cells were evaluated. rhBMP-7 is strongly mitogenic for MC3T3-E1cells, showing dose-dependent induction of ALP activity and osteocalcin production. Thus our expression system provides a con-venient source of rhBMPs for in vitro or in vivo study.. KCI Citation Count: 7