Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells.
作者： Kentaro Yamada , Masaharu Takahashi , Yu Hoshino , Hideyuki Takahashi , Koji Ichiyama
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摘要: A full-length infectious cDNA clone (pJE03-1760F/wt) of a genotype 3 hepatitis E virus (HEV) (strain JE03-1760F) obtained from faecal specimen was constructed in this study. Upon transfection the capped vitro transcripts pJE03-1760F/wt into PLC/PRF/5 cells, viral RNA levels culture supernatant started to increase on day 6 post-transfection (p.t.) and reached 107 copies ml−1 28 p.t. Detection increasing numbers cells with ORF2 protein expression by immunofluorescence assay at 5, 7, 11 15 days indicated spread HEV infection cell culture. When cDNA-derived inoculated or A549 it grew as efficiently faeces-derived both reaching 106 copies ml−1 30 days post-inoculation. Our reverse genetics system for that is usable robust cell-culture will be useful elucidation mechanism replication functional roles proteins.
sgmjournals.org参考文章(28)
E el Ghazzawi, A Panà, A M Degener, E Renganathan, F Gamil, R Gabrieli, M Divizia, O A Kader, J Pillot, E el Sherbini, G el Sawaf, M L Stefanoni, E Saleh, Evidence of hepatitis E virus replication on cell cultures. New Microbiologica. ,vol. 22, pp. 77- 83 ,(1999)
J Meng, P Dubreuil, J Pillot, A new PCR-based seroneutralization assay in cell culture for diagnosis of hepatitis E. Journal of Clinical Microbiology. ,vol. 35, pp. 1373- 1377 ,(1997) , 10.1128/JCM.35.6.1373-1377.1997
Shipra Agrawal, Dinesh Gupta, Subrat Kumar Panda, None, The 3' end of hepatitis E virus (HEV) genome binds specifically to the viral RNA-dependent RNA polymerase (RdRp). Virology. ,vol. 282, pp. 87- 101 ,(2001) , 10.1006/VIRO.2000.0819
Masaharu Takahashi, Kentaro Yamada, Yu Hoshino, Hideyuki Takahashi, Koji Ichiyama, Toshinori Tanaka, Hiroaki Okamoto, Monoclonal antibodies raised against the ORF3 protein of hepatitis E virus (HEV) can capture HEV particles in culture supernatant and serum but not those in feces Archives of Virology. ,vol. 153, pp. 1703- 1713 ,(2008) , 10.1007/S00705-008-0179-6
E. V. Koonin, A. E. Gorbalenya, M. A. Purdy, M. N. Rozanov, G. R. Reyes, D. W. Bradley, Computer-assisted assignment of functional domains in the nonstructural polyprotein of hepatitis E virus: delineation of an additional group of positive-strand RNA plant and animal viruses Proceedings of the National Academy of Sciences of the United States of America. ,vol. 89, pp. 8259- 8263 ,(1992) , 10.1073/PNAS.89.17.8259
Felipe R. Lorenzo, Toshinori Tanaka, Hideyuki Takahashi, Koji Ichiyama, Yu Hoshino, Kentaro Yamada, Jun Inoue, Masaharu Takahashi, Hiroaki Okamoto, Mutational events during the primary propagation and consecutive passages of hepatitis E virus strain JE03-1760F in cell culture. Virus Research. ,vol. 137, pp. 86- 96 ,(2008) , 10.1016/J.VIRUSRES.2008.06.005
Shaojing Wei, Peter Walsh, Rutong Huang, Shing Shun Tony To, 93G, a novel sporadic strain of hepatitis E virus in South China isolated by cell culture Journal of Medical Virology. ,vol. 61, pp. 311- 318 ,(2000) , 10.1002/1096-9071(200007)61:3<311::AID-JMV5>3.0.CO;2-H
Albert W. Tam, Matthew M. Smith, Martha E. Guerra, Chiao-Chain Huang, Daniel W. Bradley, Kirk E. Fry, Gregory R. Reyes, Hepatitis E virus (HEV): molecular cloning and sequencing of the full-length viral genome. Virology. ,vol. 185, pp. 120- 131 ,(1991) , 10.1016/0042-6822(91)90760-9
Hiroaki Okamoto, Genetic variability and evolution of hepatitis E virus. Virus Research. ,vol. 127, pp. 216- 228 ,(2007) , 10.1016/J.VIRUSRES.2007.02.002
Judith Graff, Yi-Hua Zhou, Udana Torian, Hanh Nguyen, Marisa St. Claire, Claro Yu, Robert H. Purcell, Suzanne U. Emerson, Mutations within Potential Glycosylation Sites in the Capsid Protein of Hepatitis E Virus Prevent the Formation of Infectious Virus Particles Journal of Virology. ,vol. 82, pp. 1185- 1194 ,(2008) , 10.1128/JVI.01219-07