HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplementary Table Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Google Scholar Articles by Yamada, K. Articles by Okamoto, H. PubMed PubMed Citation Articles by Yamada, K. Articles by Okamoto, H. Agricola Articles by Yamada, K. Articles by Okamoto, H.

Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells

Kentaro Yamada

Division of Virology, Department of Infection and Immunity, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke-Shi, Tochigi-Ken 329-0498, Japan

Correspondence
Hiroaki Okamoto
hokamoto{at}jichi.ac.jp

A full-length infectious cDNA clone (pJE03-1760F/wt) of a genotype 3 hepatitis E virus (HEV) (strain JE03-1760F) obtained from a faecal specimen was constructed in this study. Upon transfection of the capped in vitro transcripts of pJE03-1760F/wt into PLC/PRF/5 cells, the viral RNA levels in the culture supernatant started to increase on day 6 post-transfection (p.t.) and reached 10⁷ copies ml^–1 on day 28 p.t. Detection of increasing numbers of cells with ORF2 protein expression by immunofluorescence assay at 5, 7, 11 and 15 days p.t. indicated the spread of HEV infection in cell culture. When the cDNA-derived virus in culture supernatant was inoculated into PLC/PRF/5 or A549 cells, it grew as efficiently as the faeces-derived virus in both cells, reaching 10⁶ copies ml^–1 at 30 days post-inoculation. Our reverse genetics system for HEV that is usable in a robust cell-culture system will be useful for elucidation of the mechanism of HEV replication and functional roles of HEV proteins.

Present address: Division of Infectious Diseases, Department of Social and Environmental Medicine, Institute of Scientific Research, Oita University, Oita 879-5593, Japan.

The nucleotide sequences reported in this study have been assigned DDBJ/EMBL/GenBank accession numbers AB437316 (pJE03-1760F/wt) and AB437319 (pJE03-1760F/ORF1).

This article has been cited by other articles:

				T. Tanaka, M. Takahashi, H. Takahashi, K. Ichiyama, Y. Hoshino, S. Nagashima, H. Mizuo, and H. Okamoto Development and Characterization of a Genotype 4 Hepatitis E Virus Cell Culture System Using a HE-JF5/15F Strain Recovered from a Fulminant Hepatitis Patient J. Clin. Microbiol., June 1, 2009; 47(6): 1906 - 1910. [Abstract] [Full Text] [PDF]