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Journal of Virology, July 2008, p. 6419-6426, Vol. 82, No. 13
0022-538X/08/$08.00+0     doi:10.1128/JVI.00514-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

A Seven-Segmented Influenza A Virus Expressing the Influenza C Virus Glycoprotein HEF{triangledown}

Qinshan Gao,1 Edward W. A. Brydon,1,{dagger} and Peter Palese1,2*

Departments of Microbiology,1 Medicine, Mount Sinai School of Medicine, New York, New York 100292

Received 7 March 2008/ Accepted 21 April 2008

Influenza viruses are classified into three types: A, B, and C. The genomes of A- and B-type influenza viruses consist of eight RNA segments, whereas influenza C viruses only have seven RNAs. Both A and B influenza viruses contain two major surface glycoproteins: the hemagglutinin (HA) and the neuraminidase (NA). Influenza C viruses have only one major surface glycoprotein, HEF (hemagglutinin-esterase fusion). By using reverse genetics, we generated two seven-segmented chimeric influenza viruses. Each possesses six RNA segments from influenza virus A/Puerto Rico/8/34 (PB2, PB1, PA, NP, M, and NS); the seventh RNA segment encodes either the influenza virus C/Johannesburg/1/66 HEF full-length protein or a chimeric protein HEF-Ecto, which consists of the HEF ectodomain and the HA transmembrane and cytoplasmic regions. To facilitate packaging of the heterologous segment, both the HEF and HEF-Ecto coding regions are flanked by HA packaging sequences. When introduced as an eighth segment with the NA packaging sequences, both viruses are able to stably express a green fluorescent protein (GFP) gene, indicating a potential use for these viruses as vaccine vectors to carry foreign antigens. Finally, we show that incorporation of a GFP RNA segment enhances the growth of seven-segmented viruses, indicating that efficient influenza A viral RNA packaging requires the presence of eight RNA segments. These results support a selective mechanism of viral RNA recruitment to the budding site.

* Corresponding author. Mailing address: Department of Microbiology, Mount Sinai School of Medicine, Box 1124, One Gustave L. Levy Place, New York, NY 10029. Phone: (212) 241-7318. Fax: (212) 534-1684. E-mail: peter.palese{at}mssm.edu

{triangledown} Published ahead of print on 30 April 2008.

{dagger} Present address: Multiple Sclerosis Research Center of New York, 521 West 57th Street, 4th Floor, New York, NY 10019.

Journal of Virology, July 2008, p. 6419-6426, Vol. 82, No. 13
0022-538X/08/$08.00+0     doi:10.1128/JVI.00514-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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