Cloning and expression of Tachypleus tridentatus factor C
文献类型：期刊论文
| 作者 | Wang, DN; Liu, JW; Chen, L; Wang, L; Yang, GZ; Wu, XF; Zhang, WJ |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2002 |
| 卷号 | 34期号:1页码:77-82 |
| 关键词 | Tachypleus tridentatus factor C cloning expression bacteriostatic activity |
| 通讯作者 | , |
| 英文摘要 | Factor C is an endotoxin-sensitive, intracellular serine protease zymogen which initiates the coagulation cascade system in the horseshoe crab hemolymph. The special lipopolysaccharide (LPS) binding activation of FC makes it a potential drug for anti-LPS treatment and has a high commercial value. Based on the sequence of reported FC from Japan horseshoe crab, two pairs of primers were designed. The total RNA was extracted from amebocytes of Chinese Tachypleus tridentatus and the cDNA was separated into two part: and were cloned using RT-PCR, respectively. FCs from different geographical areas showed high homology in sequence. The whole FC cDNA was cloned into pET-28a ( + ) containing T7 promoter and recombinant expression plasmid pET-FC was constructed. The recombinant plasmid was transformed into E. coli BL21 (DE3). Recombinant FC was expressed as inclusion body when the expression strain was induced with 1 mmol/L IPTG. Refolded recombinant FC was confirmed to be active by bacteriostatic assay in vitro. The result. of Western blot also suggested the recombinant FC may be able to cleave itself partly and produced an extra immunoblot band. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | SERINE-PROTEASE ZYMOGEN ; HORSESHOE-CRAB HEMOCYTES ; PURIFICATION ; ACTIVATION |
| 收录类别 | SCI |
| 语种 | 中文 |
| WOS记录号 | WOS:000173426700015 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2425]  |
| 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 | Wang, DN,Liu, JW,Chen, L,et al. Cloning and expression of Tachypleus tridentatus factor C[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2002,34(1):77-82. |
| APA | Wang, DN.,Liu, JW.,Chen, L.,Wang, L.,Yang, GZ.,...&Zhang, WJ.(2002).Cloning and expression of Tachypleus tridentatus factor C.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,34(1),77-82. |
| MLA | Wang, DN,et al."Cloning and expression of Tachypleus tridentatus factor C".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 34.1(2002):77-82. |
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