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Journal of Virology, January 2000, p. 547-551, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Plasmid-Driven Formation of Influenza
Virus-Like Particles
Gabriele
Neumann,1
Tokiko
Watanabe,1,2 and
Yoshihiro
Kawaoka1,*
Department of Pathobiological Sciences,
School of Veterinary Medicine, University of Wisconsin
Madison,
Madison, Wisconsin 53706,1 and
Laboratory of Microbiology, Department of Disease Control,
Graduate School of Veterinary Medicine, Hokkaido University, Sapporo
060-0818, Japan2
Received 24 June 1999/Accepted 16 September 1999
We established a plasmid-based system for generating infectious
influenza virus-like particles entirely from cloned cDNAs. Human
embryonic kidney cells (293T) were transfected with plasmids encoding
the influenza A virus structural proteins and with a plasmid encoding
an influenza virus-like viral RNA (vRNA) which contained an antisense
copy of the cDNA for green fluorescence protein (GFP) flanked by an RNA
polymerase I promoter and terminator. Intracellular transcription of
the latter construct by RNA polymerase I generated GFP vRNA that was
packaged into influenza virus-like particles. This system, which
produced more than 104 infectious particles per ml of
supernatant, would be useful in studies of influenza virus replication
and particle formation. It might also benefit efforts in vaccine
production and in the development of improved gene therapy vectors.
*
Corresponding author. Mailing address: Department of
Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin
Madison, 2015 Linden Dr. West, Madison, WI 53706. Phone: (608) 265-4925. Fax: (608) 265-5622. E-mail:
kawaokay{at}svm.vetmed.wisc.edu.
Journal of Virology, January 2000, p. 547-551, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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