High-level expression of the C-terminal hydrophobic region of HCV E2 protein ectodomain in E-coli
文献类型：期刊论文
| 作者 | Liu, J; Kong, YY; Zhu, LX; Wang, Y; Li, GD |
| 刊名 | VIRUS GENES
 |
| 出版日期 | 2002 |
| 卷号 | 25期号:1页码:42137 |
| 关键词 | E2 protein expression in E. coli hepatitis C virus hexa-histidine-tagged protein hydrophobic region purification |
| 通讯作者 | Wang, Y (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China., |
| 英文摘要 | High-level expression of hepatitis C virus (HCV) E2 protein fragments encompassing its C-terminal hydrophobic region (downstream of as 661) in Escherichia coli has been proven difficult. The extreme hydrophobicity of this region has been suspected to be detrimental to the host. In this work, we found that the C-terminal region (downstream of as 565) of E2 ectodomain interfered with full-length expression of E2 fragments in E. coli. Nonetheless, when the central region (aa 484-622) of E2 was deleted, full-length protein was efficiently produced. C-terminal region as 567-700 could not be efficiently expressed individually or as mouse dihydrofolate reductase (DHFR) fusion protein. However, a mutant that emerged in the cloning process was able to express full-length DHFR fusion protein. Sequencing analysis reveals the mutation to be a short frame-shift around the fusion junction, altering as 568-571 of E2. C-terminal region of E2 ectodomain (aa 567-730) carrying this mutation was successfully expressed as hexa-histidine-tagged protein to a high level. The protein was highly insoluble and was purified under denaturing conditions. The purified protein displayed HCV E2-specific antigenicity in Western blot and specific rabbit antiserum was raised against it. These results demonstrate that hydrophobicity of the C-terminal region of E2 ectodomain is not harmful to E. coli host and has no dominative adverse effect on its bacterial expression. Other nucleotide and/or amino acid sequence properties seem to play a more important role. This finding opens up new possibilities for the development of novel bacterially-derived E2 proteins for research and clinical applications. |
| 学科主题 | Genetics & Heredity; Virology |
| 类目[WOS] | Genetics & Heredity ; Virology |
| 关键词[WOS] | ESCHERICHIA-COLI ; LIVER-DISEASE ; ANTI-HCV ; VIRUS ; GLYCOPROTEIN ; INFECTION ; CELLS ; IDENTIFICATION ; ANTIBODIES ; PRODUCTS |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000177582700001 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2407]  |
| 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 | Liu, J,Kong, YY,Zhu, LX,et al. High-level expression of the C-terminal hydrophobic region of HCV E2 protein ectodomain in E-coli[J]. VIRUS GENES,2002,25(1):42137. |
| APA | Liu, J,Kong, YY,Zhu, LX,Wang, Y,&Li, GD.(2002).High-level expression of the C-terminal hydrophobic region of HCV E2 protein ectodomain in E-coli.VIRUS GENES,25(1),42137. |
| MLA | Liu, J,et al."High-level expression of the C-terminal hydrophobic region of HCV E2 protein ectodomain in E-coli".VIRUS GENES 25.1(2002):42137. |
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