Secretory expression of different C-terminal truncated HCV E1 proteins in mammalian cells and characterization of the expressed products
文献类型：期刊论文
| 作者 | Zhu, J; Kong, YY; Liu, J; Zhang, ZC; Wang, Y; Li, GD |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2001 |
| 卷号 | 33期号:6页码:634-640 |
| 关键词 | HCV E1 HBV preS1 double antigenicity secretary expression glycosylation |
| 通讯作者 | , |
| 英文摘要 | Three fragments of the HCV envelope 1 (EI) with different C-terminal truncation at aa310, aa325, aa340 were cloned into the mammalian expression vector pSecTagB. An epitope in the hepatitis B surface antigen, preS1 (21-47), were genetically engineered onto the N-terminus of the recombinant protein and used as an affinity tag for detection and purification. The resulting pSec-preS1-E1t310, pSec-preS1-E1t325 and pSec-preS1-E1t340 were transiently expressed in the HeLa cells and the antigenicity, secretory efficiency and glycosylation type of the recombinant El proteins were compared. All of the three recombinant proteins could be detected by both preS1 monoclonal antibody and El polyclonal antiserum. The expression products were secreted and highly mannose-type glycosylated, with S1E1t325 being secreted, indicating the influence of the hydrophobic regions on the secretion of the El protein. Three CHO cell lines expressing the proteins, S1E1t310, S1E1t325 and S1E1t340, were established and the CHO/pSecS1E1t325 was chosen for further study. The secreted S1E1t325 could be enriched from cell culture medium by the preS1 antibody-coupled Sepharose. The glycosylation analysis indicated the lack of complex glycogen even after the El was secreted via Golgi complexes. The established stable cell lines and anti-preS1 affinity method could be utilized to enrich and purify the HCV El expressed in mammalian cells, and may be used for further characterization of this protein. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | ESCHERICHIA-COLI ; STRUCTURAL PROTEINS ; GLYCOPROTEIN E1 ; INSECT CELLS ; VIRUS HCV ; REGION ; INFECTION ; E2 |
| 收录类别 | SCI |
| 语种 | 中文 |
| WOS记录号 | WOS:000172362600007 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2687]  |
| 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 | Zhu, J,Kong, YY,Liu, J,et al. Secretory expression of different C-terminal truncated HCV E1 proteins in mammalian cells and characterization of the expressed products[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2001,33(6):634-640. |
| APA | Zhu, J,Kong, YY,Liu, J,Zhang, ZC,Wang, Y,&Li, GD.(2001).Secretory expression of different C-terminal truncated HCV E1 proteins in mammalian cells and characterization of the expressed products.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,33(6),634-640. |
| MLA | Zhu, J,et al."Secretory expression of different C-terminal truncated HCV E1 proteins in mammalian cells and characterization of the expressed products".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 33.6(2001):634-640. |
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