%0 Journal Article
%T Expression, Characterization and Biological Activity Analysis of Recombinant Human Bone Morphogenetic Protein 2 in CHO Cells<br>重组人骨形态发生蛋白2在CHO细胞中的表达、鉴定及活性分析
%A ZHANG Dao-Yong
%A YANG Shuang
%A L
%A <br>张道永
%A 杨爽
%A 吕树军
%A 闫继东
%A 朱天慧
%J 生物工程学报
%D 2006
%I 
%X Bone morphogenetic protein 2(BMP-2) is a member of the of BMPs family, its osteoinductive capacity has already been demonstrated. We tried to express hBMP-2 in CHO cell. In this study, we inserted hBMP-2 cDNA into vector pCDNA3.1(+) to construct hBMP-2 eukaryotic expression vector pCDNA3.1(+)-hBMP-2. Recombinant Chinese hamster ovary (rCHO) cell line expressing high-level recombinant human bone morphogenetic protein 2(rhBMP-2) was constructed by co-transfecting the expression vectors pCDNA3.1(+)-hBMP-2 and plasmid pSV2-dhfr into dihydrofolate reductase (dhfr)-deficient CHO cells and the subsequent gene amplification in medium containing stepwise increments in methotrexate level such as 0.1 and 1 micromol/L. Western blot analyses showed a specific band of about 18 kD in reduced sample lane and a specific band of about 32 kD in non-reduced sample lane, this indicated that rCHO cells secret rhBMP-2 as a homodimeric glycoprotein form. Finally, we obtained a single clone cell strain expressing a high level (7.83 microg/24 h/10(6) cells) of rhBMP-2 tested by ELISA. Biological activity of rhBMP-2 was tested by the induction of alkaline phosphatase(ALP) activity in C2C12 cells. We treated C2C12 with different concentration of rhBMP-2 condition medium(CM) for 5d. The results showed that the rhBMP-2 could significantly increase the ALP activity of C2C12.
%K human bone morphogenetic protein 2
%K CHO
%K co-transfect
%K C2C12
%K ALP<br>人骨形态发生蛋白2
%K CHO
%K 共转染
%K C2C12
%K 碱性磷酸酶
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=78CCA7D5CA9DF25A&yid=37904DC365DD7266&vid=BC12EA701C895178&iid=B31275AF3241DB2D&sid=20154CF366EC08B1&eid=8115E88DD41C4B46&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=2&reference_num=10