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Production of bone morphogenetic protein-7 using pET expression system

Authors
 Lee Dong Hee  ;  Baek Hyun Sook  ;  Lee Mi Hee  ;  Park Jong-Chul 
Citation
 CURRENT APPLIED PHYSICS, Vol.5(5) : 422-425, 2005 
Journal Title
CURRENT APPLIED PHYSICS
ISSN
 1567-1739 
Issue Date
2005
MeSH
Bone morphogenetic protein ; E. coli ; Inclusion body ; n-Cethyltrimethylammonium chloride ; Refolding ; Osteoblast
Keywords
Bone morphogenetic protein ; E. coli ; Inclusion body ; n-Cethyltrimethylammonium chloride ; Refolding ; Osteoblast
Abstract
The polypeptide representing the mature part of human bone morphogenetic protein-7 (BMP-7) was efficiently expressed in E. coli BL21 (DE3). The rhBMP-7 is a disulfide-bonded homodimeric protein with an apparent MW of 29,000 as shown by the SDS-PAGE and gel chromatography. rhBMP-7 stimulates ALP specific activity in a dose-dependent manner. In a kinetic experiment, ALP activity in both the rhBMP-7 (100 ng/ml)-treated and control cultures increases gradually, and activity in the rhBMP-7-treated culture is consistently higher throughout the culture period. We also quantitated the specific osteoblastic marker osteocalcin in rhBMP-7-treated culture medium. When serum-starved MC3T3-E1 cells are treated with medium containing various concentrations of rhBMP-7, the production of osteocalcin is increased about 7.2-fold in a dose-dependent manner up to 320 ng rhBMP-7/ml. In summary, we established an E. coli expression system for a high level of rhBMP-7 production and inductive effects of osteogenic differentiation markers by rhBMP-7 in MC3T3-E1 cells were evaluated. rhBMP-7 is strongly mitogenic for MC3T3-E1 cells, showing dose-dependent induction of ALP activity and osteocalcin production. Thus our expression system provides a convenient source of rhBMPs for in vitro or in vivo study.
Full Text
http://www.sciencedirect.com/science/article/pii/S1567173905000076
DOI
10.1016/j.cap.2005.01.003
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Medical Engineering (의학공학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Yonsei Biomedical Research Center (연세의생명연구원) > 1. Journal Papers
Yonsei Authors
Park, Jong Chul(박종철) ORCID logo https://orcid.org/0000-0003-0083-5991
Baek, Hyun Sook(백현숙)
Lee, Dong Hee(이동희)
Lee, Mi Hee(이미희) ORCID logo https://orcid.org/0000-0002-9630-7044
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/147496
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