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Clone and expression of human osteoprotegerin gene
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 22
- DOI:
- 10.3321/j.issn:1000-2790.2002.22.005
- Key Word:
- 破骨细胞;抑制因子免疫;逆转录聚合酶链反应;基因表达
Abstract： 目的克隆、表达和鉴定人破骨细胞抑制因子(OPG). 方法从培养的人胚肺成纤维细胞(WI-38)中提取总RNA,经RT-PCR获得人破骨细胞抑制因子基因. 将该基因克隆到pGEM3zf-载体中,测序鉴定. 继而将OPG基因插入TNFHis融合表达载体中,42℃诱导表达4～5 h,做SDS-PAGE分析. 以免疫印迹法鉴定TNFHis OPG融合蛋白的表达. 结果 DNA测序证明,获得了OPG基因,其序列与国外文献报道不完全一致. SDS-PAGE分析表明,TNFHisOPG融合蛋白获得表达,其Mr为7.1×104 ku,表达量约占菌体总蛋白的10%. 免疫印迹法鉴定显示,该融合蛋白可与抗TNF-α单抗产生阳性反应. 结论 OPG基因的克隆和表达均获得了成功.
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