High-level expression and secretion of recombinant mouse endostatin by Escherichia coli
文献类型：期刊论文
| 作者 | Xu, R; Du, P; Fan, JJ; Zhang, Q; Li, TP; Gan, RB |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2002 |
| 卷号 | 24期号:3页码:453-459 |
| 关键词 | endostatin secretory expression angiogenesis G(1) arrest |
| 通讯作者 | Xu, R (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, POB 55,320 Yue Yang Rd, Shanghai 200031, Peoples R China., |
| 英文摘要 | The expression of murine endostatin was achieved by placing its gene downstream of an alkaline phosphatase gene (phoA) promoter. To ensure proper folding and secretion of the recombinant protein, the mouse endostatin was fused with alkaline phosphatase signal peptide. SDS/polyacrylamide gel electrophoresis analysis of the culture medium of recombinant Escherichia coli cells revealed that endostatin was efficiently secreted. The signal peptide was efficiently cleaved during secretion as demonstrated by N-terminal amino acid sequencing. The maximum yield of secreted endostatin during fermentation was 40 mg/liter. Up to 28 mg of endostatin was purified from 1 liter of cell culture broth. The biological activity of recombinant protein was tested in a bovine aortic endothelial (BAE) cell proliferation assay. The recombinant endostatin inhibited the growth of BAE cells stimulated by basi. Le fibroblast growth factor, and its ED50 was comparable to that from a previous report. Flow cytometric measurements of BAE cells cultivated in medium with endostatin demonstrated a cell cycle arrest mainly in the G(0)/G(1) phase and a decrease in the S phase. (C) 2002 Elsevier Science (USA). |
| 学科主题 | Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology |
| 类目[WOS] | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 关键词[WOS] | ANGIOGENESIS INHIBITOR ENDOSTATIN ; CELL APOPTOSIS ; GROWTH-FACTOR ; TUMOR-GROWTH ; GENE |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000174963400016 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2469]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Xu, R,Du, P,Fan, JJ,et al. High-level expression and secretion of recombinant mouse endostatin by Escherichia coli[J]. PROTEIN EXPRESSION AND PURIFICATION,2002,24(3):453-459. |
| APA | Xu, R,Du, P,Fan, JJ,Zhang, Q,Li, TP,&Gan, RB.(2002).High-level expression and secretion of recombinant mouse endostatin by Escherichia coli.PROTEIN EXPRESSION AND PURIFICATION,24(3),453-459. |
| MLA | Xu, R,et al."High-level expression and secretion of recombinant mouse endostatin by Escherichia coli".PROTEIN EXPRESSION AND PURIFICATION 24.3(2002):453-459. |
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