Investigation of norovirus replication in a human cell line

Archives of Virology
Kazuhiko KatayamaN Takeda

Abstract

Noroviruses (NoVs) belong to the genus Norovirus and are members of the family Caliciviridae. NoVs are the dominant cause of outbreaks of gastroenteritis, but progress in understanding the molecular characteristics of NoV and its replication strategies have been hampered by the lack of a cell culture system or a practical animal model, except for murine NoVs. To elucidate the transcription and replication of the NoV genome, a complete genome of a human NoV genogroup II strain was cloned downstream of a T7 RNA polymerase promoter and expressed in human embryonic kidney (HEK) 293T/17 cells using a T7 vaccinia virus expression system. Bands for a 7.6-kb negative-strand RNA, a 7.6-kb positive-strand genomic RNA, and a 2.6-kb positive-strand subgenomic-like RNA were found in the infected cells. However, recombinant capsid protein (rVP1) and rVP2 were not detected by Western blotting. When a construct containing VP1 and VP2 genes was co-transfected with a full-length construct, the expression of virus-like particles (VLPs) with a buoyant density of 1.271 g/cm3 was observed. We also observed round particles, 20 to 80 nm in diameter, with a buoyant density of 1.318 g/cm3. Our results indicated that NoV RNA was incorporated into the hea...Continue Reading

References

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Citations

Nov 24, 2006·Journal of Virology·Stephen W B FullertonJacques Rohayem
May 16, 2006·Journal of Virology·Christiane E WobusHerbert W Virgin
Oct 14, 2011·Journal of Virology·Chennareddy V Subba-ReddyC Cheng Kao
Jun 15, 2007·Virology Journal·Karen KearneyJohn G Morgan
Sep 21, 2010·Uirusu·Hironori Sato
Oct 2, 2008·Emerging Infectious Diseases·Robert L AtmarDavid Y Graham
Sep 7, 2014·Proceedings of the National Academy of Sciences of the United States of America·Kazuhiko KatayamaMary K Estes
Sep 15, 2007·Journal of Virology·Susana GuixMary K Estes
Jan 19, 2016·Archives of Virology·Sangdo HaJinjong Myoung
Oct 8, 2008·Immunological Reviews·Eric F DonaldsonRalph S Baric
Jan 6, 2011·Microbiology and Immunology·Tomoichiro OkaKazuhiko Katayama
Dec 3, 2014·Trends in Microbiology·Stefan Taube, Christiane E Wobus
Mar 1, 2010·Viruses·Stephanie M Karst
Jan 9, 2015·Clinical Microbiology Reviews·Tomoichiro OkaLinda J Saif
Feb 15, 2019·Viruses·Kyle V Todd, Ralph A Tripp
Jan 23, 2017·Water Research·Sorina E EftimSharon P Nappier

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