High-level production of a functional recombinant hepatitis b virus polymerase in insect cells with a baculovirus expression system
文献类型：期刊论文
| 作者 | Wang Xiaoyan1; Gao Linlin1; Deng Fei2; Zhang Yanfang2; Li Yan1; Lin Jusheng1 |
| 刊名 | Journal of huazhong university of science and technology-medical sciences
 |
| 出版日期 | 2007-06-01 |
| 卷号 | 27期号:3页码:269-273 |
| 关键词 | Hepatitis b virus Polymerase Baculovirus Insect cell |
| ISSN号 | 1672-0733 |
| DOI | 10.1007/s11596-007-0313-9 |
| 通讯作者 | Lin jusheng(linjusheng2001@163.com) |
| 英文摘要 | Hbv polymerase has intrinsic rna-dependent reverse transcriptase, dna-dependent dna polymerase as well as rnaseh activity. analysis of hbv polymerase has been hampered for many years due to the inability to express functional enzyme in a recombinant system. to obtain active polymerase at a high level, we have taken advantage of baculovirus expression system. the gene of hbv polymerase was amplified by pcr and cloned into pfastbac dual to construct the recombinant plasmid pfastbac dual-pol. the recombinant donor plasmid, pfastbac dual-pol, was constructed by inserting hbv polymerase gene into ecori and psti sites controlled by polyhedrin promoter. the recombinant donor plasmid was transformed into dh10bac competent cells for transposition. recombinant bacmid was constructed by inserting of the mini-tn7 element from the donor plasmid into the mini-atttn7 attachment site on the bacmid. the recombinant bacmid dna was isolated and transfected into the sf9 cells to produce the recombinant virus, and healthy insect sf9 cells were infected with the recombinant virus containing hbv polymerse-gene to express the target protein. hbv polymerse expressed in insect cells was analyzed by sds-page. pcr results showed recombinant donor plasmid, pfastbac dual-pol, was constructed successfully. the recombinant hepatitis b virus polymerase was expressed in insect cells at high level. the recombinant hepatitis b virus polymerase should facilitate the analysis of hbv polymerase biological characteristics, allow the investigation for new anti-hbv drugs specifically blocking hbv polymerase. |
| WOS关键词 | P-GENE-PRODUCT ; REVERSE TRANSCRIPTION ; PROTEIN ; GENOME ; DNA ; REPLICATION ; INITIATION ; BIOLOGY ; DOMAIN |
| WOS研究方向 | Biochemistry & Molecular Biology |
| WOS类目 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000257074000013 |
| 出版者 | SPRINGER |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2375371 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Lin Jusheng |
| 作者单位 | 1.Huazhong Univ Sci & Technol, Tongii Med Coll, Tongji Hosp, Inst Liver Dis, Wuhan 430030, Peoples R China 2.Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430030, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang Xiaoyan,Gao Linlin,Deng Fei,et al. High-level production of a functional recombinant hepatitis b virus polymerase in insect cells with a baculovirus expression system[J]. Journal of huazhong university of science and technology-medical sciences,2007,27(3):269-273. |
| APA | Wang Xiaoyan,Gao Linlin,Deng Fei,Zhang Yanfang,Li Yan,&Lin Jusheng.(2007).High-level production of a functional recombinant hepatitis b virus polymerase in insect cells with a baculovirus expression system.Journal of huazhong university of science and technology-medical sciences,27(3),269-273. |
| MLA | Wang Xiaoyan,et al."High-level production of a functional recombinant hepatitis b virus polymerase in insect cells with a baculovirus expression system".Journal of huazhong university of science and technology-medical sciences 27.3(2007):269-273. |
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来源：武汉病毒研究所
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