[Expression of recombinant human BMP6 in CHO cells by fused to the signal peptide and propeptide of another homologue protein].
BMP6 is a member of TGF-beta superfamily, represent more effective osteogenic activity. Two recombinant plasmids were constructed to expression rhBMP6 in mammalian cells, one contained the cDNA encoding the signal peptide, propeptide and mature peptide of human BMP6, wich was named pcDNA-BMP6, the other contained the recombinant DNA encoding the signal peptide, propeptide of human BMP2 and the mature peptide of BMP6, which was named pcDNA-BMP2/6. Transient expression in Cos7 cells demonstrated that the pcDNA-BMP2/6 produced more rhBMP6 than pcDNA-BMP6. For stable expression, the CHO-dhfr- cells were transfected with pcDNA-BMP2/6 and pSV2-dhfr, then screened by G418 and treated with MTX for targeting gene amplification. The partially purified rhBMP6 by heparin affinity chromatography was shown to possess bone induction activity tested by the induction of alkaline phosphatase activity in C2C12 cells.
Yan
Ji-Dong
JD
Laboratory of Molecular Genetics, College of Medicine, Nankai University, Tianjin 300071, China.
Yang
Shuang
S
Lü
Shu-Jun
SJ
Lei
Rong-Yue
RY
Zhu
Tian-Hui
TH
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D055418:Bone Morphogenetic Protein 6 / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D006801:Humans; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D011498:Protein Precursors / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D055418:Bone Morphogenetic Protein 6; D011498:Protein Precursors; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase