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重组人成骨生长肽的表达、纯化和活性的研究
Expression, Purification and Activity of Recombinant Human Osteogenic Growth Peptide (rhOGP)
【摘要】 将人工合成的人成骨生长肽基因 (hOGPgene) ,与质粒pTYB2 重组 ,转化大肠杆菌E .coliBL2 1 (DE3) .经异丙基硫代 β D 半乳糖苷 (IPTG)诱导 ,融合蛋白在E .coli中得到表达 .菌体经超声破碎 ,一步亲和层析纯化得到重组人成骨生长肽 (recombinanthumanosteogenicgrowthpeptide ,rhOGP) .体外实验证明 :rhOGP对成纤维细胞 (NIH3T3)的增殖有促进作用 .体内实验证明 :rhOGP加速兔移植骨成活 ,使血清中碱性磷酸酶活性增高 ,骨钙素增高 .重组人成骨生长肽促进成骨 ,有可能成为治疗骨移植及骨折愈合的有效药物
【Abstract】 Synthetic human gene of osteogenic growth peptide (hOGP gene) was inserted into the plasmid pTYB 2. This recombinant plasmid was transformed into E.coli strain BL21(DE3). Transformants were induced by IPTG, and then expressed. After purification by only one step affinity chromatography with ultrasonification of E.coli, the pure rhOGP has been obtained. This can stimulate the proliferation of fibroblast NIH3T3 in vitro. Moreover, It can enhance the healing of bone grafting of rabbit, increase the activity of alkaline phosphatase and promote the level of osteocalcin in vivo. Therefore, The rhOGP can promote osteogenesis, and rhOGP has potential roles in accelerating the healing of bone grafting as well as fractures.
【Key words】 rhOGP; fusion protein; affinity chromatography; activity; ostogenesis;
- 【文献出处】 中国生物化学与分子生物学报 ,Chinese Journal of Biochemistry and Molecular Biology , 编辑部邮箱 ,2004年04期
- 【分类号】Q516
- 【被引频次】17
- 【下载频次】224