科研成果
| 论文编号: | DOI:10.1042/BA20070225 |
| 第一作者所在部门: | 天然产物研究中心 |
| 论文题目: | Novel preparation protocol for the expression and purification of recombinant staphylokinase |
| 作者: | Dongsheng Ren,Dehua Li |
| 全部作者: | Dongsheng Ren,Dehua Li,Weihua Yang,Yun Li,Xinghua Gou,Bo Liang,Bogang Li,Qiaqing Wu* |
| 论文出处: | |
| 刊物名称: | Biotechnology and Applied Biochemistry |
| 年: | 2008 |
| 卷: | 51 |
| 期: | 1 |
| 页: | 9-13 |
| 联系作者: | 吴洽庆 |
| 收录类别: | |
| 影响因子: | 1.288 |
| 摘要 : | Staphylokinase (Sak), produced by lysogenic strains of Staphylococcus aureus, can convert plasminogen into its proteolytic form, plasmin, and thus is widely used to dissolve pathological clots in clinical applications. In the present paper, we report a novel approach to produce r-Sak (recombinant Sak)using an engineered Escherichia coli expression system. The expression plasmid was constructed by placing the Sak gene into the expression vector pET32(a), resulting in the expression of 35% fusion protein. Subsequently, a rapid and simple chromatographic procedure was developed for the large-scale purification of therapeutic-grade r-Sak from E. coli, which includes Ni2+-affinity chromatography,ultrafiltration and Q-Sepharose Fast Flow chromatography.This method led to the production of highly pure r-Sak (>99 %), according to SDS/PAGE and HPLC analysis. |
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