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	Baculovirus mediated production of infectious hepatitis c virus in human hepatoma cells stably expressing t7 rna polymerase
	Yao, Xiangjie1,2; Han, Qingxia1,2; Song, Jianhua1; Liang, Changyong1; Wakita, Takaji3; Yang, Rongge1; Chen, Xinwen1
刊名	Molecular biotechnology
	2008-10-01
卷号	40期号:2页码:186-194
关键词	Hcv Baculovirus vector Infectious particles Replication
ISSN号	1073-6085
DOI	10.1007/s12033-008-9075-2
通讯作者	Chen, xinwen(chenxw@pentium.whiov.ac.cn)
英文摘要	Although much has been learned about hepatitis c virus (hcv), research progress has been hindered by the lack of a suitable cell culture system supporting its replication. recently, a unique hcv strain jfh1 has been found to replicate efficiently in cell culture with production of infectious hcv (hcvcc). baculovirus vectors were found to be efficient delivery vehicles and a hbv recombinant baculovirus/hepg2 system efficiently delivered the hbv genome into hepg2 resulting in hbv replication. in this study, we developed a recombinant baculovirus expression system to generate infectious hcv particles in hepatoma cell line huh7-lunett7 by using cdna from the hcv jfh1 genotype. results show that hcv positive, negative rna strands and proteins were produced in this system. furthermore, hcv particles were produced and secreted into the culture medium. sucrose density gradient centrifugation of the culture medium revealed co-localization of hcv rna and structural proteins in the fraction with a density of 1.08-1.13 g/ml. electron microscopy (em) showed viral particles approximately 55 nm in diameter, which could be recognized by anti-hcv e2 antibodies. real-time rt-pcr detected that the level of hcv vrna in the supernatant was 10(7) copies/ml at 72 h post-transduction (hpt). in addition, the jfh1 virus produced by the recombinant baculovirus was confirmed to be infectious in vitro. in summary, this system provides a novel tool not only for the analysis of the replication and pathogenesis of hcv but also to screen for potential therapeutic targets.
WOS关键词	FULL-LENGTH ; RECOMBINANT BACULOVIRUS ; HEPG2 CELLS ; EFFICIENT REPLICATION ; SUBGENOMIC REPLICON ; MAMMALIAN-CELLS ; HCV REPLICATION ; GENE-TRANSFER ; IN-VITRO ; RT-PCR
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS类目	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	HUMANA PRESS INC
WOS记录号	WOS:000259735600009
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2375528
专题	武汉病毒研究所
通讯作者	Chen, Xinwen
作者单位	1.Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 10039, Peoples R China 3.Natl Inst Infect Dis, Dept Virol 2, Shinjuku Ku, Tokyo 1628640, Japan
推荐引用方式 GB/T 7714	Yao, Xiangjie,Han, Qingxia,Song, Jianhua,et al. Baculovirus mediated production of infectious hepatitis c virus in human hepatoma cells stably expressing t7 rna polymerase[J]. Molecular biotechnology,2008,40(2):186-194.
APA	Yao, Xiangjie.,Han, Qingxia.,Song, Jianhua.,Liang, Changyong.,Wakita, Takaji.,...&Chen, Xinwen.(2008).Baculovirus mediated production of infectious hepatitis c virus in human hepatoma cells stably expressing t7 rna polymerase.Molecular biotechnology,40(2),186-194.
MLA	Yao, Xiangjie,et al."Baculovirus mediated production of infectious hepatitis c virus in human hepatoma cells stably expressing t7 rna polymerase".Molecular biotechnology 40.2(2008):186-194.

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