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重组人成骨蛋白-1在大肠杆菌中的高效表达及复性 被引量：8

Study on high expression and renaturation of rhOP-1
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摘要 目的研究重组人成骨蛋白-1(rhOP-1)的发酵、纯化和复性。方法将构建好的重组质粒pBV221转化EscherichiacoliBL21,迅速升温至42℃诱导rhOP-1以包涵体的形式高效表达。裂菌后利用6mol·L-1尿素溶解目的蛋白,经SP-FastFlow离子交换色谱纯化后,利用cysteine/cystine对单体rhOP-1透析复性。结果rhOP-1的表达量占菌体总蛋白质量的33%,纯化后纯度可达98%,活性得到有效恢复。结论建立了rhOP-1发酵、纯化与复性的方法,获得高表达、高纯度、有活性的rhOP-1。 OBJECTIVE: To study the fermentation, purification and renaturation of recombinant human osteogenic protein-1(rhOP-1). METHODS: Recombinant plasmid pBV221 carrying rhOP-1 gene was transformed into Escherichia coli BL21. rhOP-1 was expressed from fermented E. coli by quickly raising temperature to 42 °C. A high expression was achived after inducing for 4 h. rhOP-1 was purified by SP-FF chromatography and its activity was recovery by dialysis using cysteine/cystine or GSH/GSSG to form dimers. In vitro cell activity was confirmed by method MTT and PNPP. RESULTS: The expression of rhOP-1 was 33% of total bacterial protein. The purity of rhOP-1 was about 98% and its activity was well recovered. CONCLUSION: The process of rhOP-1 fermentation, purification and renaturation was estabolished, and the highly expressed, high purity and high activity rhOP-1 could be obtained.
作者 车婧 韩金祥 王世立 梁浩
机构地区 山东省医药生物技术研究中心
出处 《中国药学杂志》 EI CAS CSCD 北大核心 2005年第13期1023-1026,共4页 Chinese Pharmaceutical Journal
基金 国家高技术研究发展计划(863)项目(2003AA223532)山东省重大攻关项目
关键词 重组人成骨蛋白 纯化 复性 Cells Chromatographic analysis Dimers Escherichia coli Fermentation Genes Proteins Purification
分类号 Q81 [生物学—生物工程]
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参考文献11

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二级参考文献47

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中国药学杂志
2005年 第13期

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