| 利用同源蛋白的信号肽和前肽序列在CHO细胞中表达重组人BMP6 |
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| 引用本文： | 闫继东,杨爽,吕树军,雷荣悦,朱天慧. 利用同源蛋白的信号肽和前肽序列在CHO细胞中表达重组人BMP6[J]. 生物工程学报, 2007, 23(3): 413-417 |
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| 作者姓名： | 闫继东 杨爽 吕树军 雷荣悦 朱天慧 |
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| 作者单位： | 南开大学医学院分子遗传实验室,天津300071 |
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| 摘 要： | BMP6属于TGF-β超家族,具有较强的骨诱导作用。主要利用BMP6自身的信号肽、前肽与成熟肽基因序列构建了表达质粒pcDNA-BMP6;同时利用BMP2的信号肽、前肽与BMP6的成熟肽基因序列构建了表达质粒pcDNA-BMP2/6。将两种质粒分别瞬时转染Cos7细胞,发现质粒pcDNA-BMP2/6表达rhBMP6的效率高于质粒pcDNA-BMP6。然后将质粒pcDNA-BMP2/6与含二氢叶酸还原酶基因(dhfr)的表达质粒共转染dhfr缺陷型的中华仓鼠卵巢(CHO)细胞,经G418筛选、氨甲蝶呤(MTX)介导目的基因扩增、亚克隆后得到表达rhBMP6成熟肽的单克隆细胞株。将表达产物rhBMP6初步纯化后,能诱导前成肌细胞系C2C12向成骨细胞方向转化,显示其具有骨诱导作用。
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| 关 键 词： | 中华仓鼠卵巢细胞 重组表达 骨诱导作用 |
| 文章编号： | 1000-3061（2007）03-0413-05 |
| 修稿时间： | 2006-10-262006-12-19 |
Expression of Recombinant Human BMP6 in CHO Cells by Fused to the Signal Peptide and Propeptide of Another Homologue Protein |
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| YAN Ji-Dong,YANG Shuang,L Shu-Jun,LEI Rong-Yue,ZHU Tian-Hui. Expression of Recombinant Human BMP6 in CHO Cells by Fused to the Signal Peptide and Propeptide of Another Homologue Protein[J]. Chinese journal of biotechnology, 2007, 23(3): 413-417 |
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| Authors: | YAN Ji-Dong YANG Shuang L Shu-Jun LEI Rong-Yue ZHU Tian-Hui |
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| Affiliation: | Laboratory of Molecular Genetics, College of Medicine, Nankai University, Tianjin 300071, China;Laboratory of Molecular Genetics, College of Medicine, Nankai University, Tianjin 300071, China;Laboratory of Molecular Genetics, College of Medicine, Nankai University, Tianjin 300071, China;Laboratory of Molecular Genetics, College of Medicine, Nankai University, Tianjin 300071, China;Laboratory of Molecular Genetics, College of Medicine, Nankai University, Tianjin 300071, China |
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| Abstract: | 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. |
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| Keywords: | BMP6 |
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