Novel preparation protocol for the expression and purification of recombinant staphylokinase
文献类型：期刊论文
| 作者 | Ren, DS ; Li, DH ; Yang, WH ; Li, Y ; Gou, XH ; Liang, B ; Li, BG ; Wu, QQ |
| 刊名 | BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
 |
| 出版日期 | 2008 |
| 卷号 | 51期号:1页码:9_13 |
| 关键词 | Escherichia coli expression Ni2+-nitrilotriacetate purification Q-Sepharose Fast Flow chromatography recombinant staphylokinase |
| ISSN号 | 0885-4513 |
| 产权排序 | 1 |
| 通讯作者 | Wu, QQ, Chinese Acad Sci, Chengdu Inst Biol, Chengdu 610041, Peoples R China. |
| 中文摘要 | 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. |
| 学科主题 | Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000259131100002 |
| 公开日期 | 2011-07-08 |
| 源URL | [http://210.75.237.14/handle/351003/18113]  |
| 专题 | 成都生物研究所_天然产物研究 |
| 推荐引用方式 GB/T 7714 | Ren, DS,Li, DH,Yang, WH,et al. Novel preparation protocol for the expression and purification of recombinant staphylokinase[J]. BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY,2008,51(1):9_13. |
| APA | Ren, DS.,Li, DH.,Yang, WH.,Li, Y.,Gou, XH.,...&Wu, QQ.(2008).Novel preparation protocol for the expression and purification of recombinant staphylokinase.BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY,51(1),9_13. |
| MLA | Ren, DS,et al."Novel preparation protocol for the expression and purification of recombinant staphylokinase".BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY 51.1(2008):9_13. |
入库方式： OAI收割
来源：成都生物研究所
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