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Journal of Virology, September 2009, p. 9296-9303, Vol. 83, No. 18
0022-538X/09/$08.00+0     doi:10.1128/JVI.00781-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

A One-Plasmid System To Generate Influenza Virus in Cultured Chicken Cells for Potential Use in Influenza Vaccine{triangledown}

Xiangmin Zhang,1 Wei Kong,1 Shamaila Ashraf,1 and Roy Curtiss III1,2*

Center for Infectious Diseases and Vaccinology, the Biodesign Institute,1 School of Life Science, Arizona State University, Tempe, Arizona 852872

Received 16 April 2009/ Accepted 1 July 2009

Influenza virus has a set of ribonucleoproteins (RNPs) consisting of viral RNAs, influenza virus polymerase subunits, and nucleoprotein. Intracellular reconstitution of the whole set of RNPs via plasmid transfection results in the generation of influenza virus. By the use of reverse genetics and dual promoters, we constructed a 23.6-kb eight-unit plasmid that contains all the required constituents to generate influenza virus in chicken cells. Our "one-plasmid" system generated higher titers of influenza virus in chicken cells than the "eight-plasmid" system, enabling a simpler approach for generating vaccine seeds. Our study identified plasmid size as a potential limiting factor affecting transfection efficiency and hence the influenza viral yield from chicken cells.

* Corresponding author. Mailing address: Center for Infectious Diseases and Vaccinology, the Biodesign Institute, Arizona State University, P.O. Box 875401, 1001 S. McAllister Avenue, Tempe, AZ 85287-5401. Phone: (480) 727-0445. Fax: (480) 727-0466. E-mail: rcurtiss{at}asu.edu

{triangledown} Published ahead of print on 8 July 2009.

Journal of Virology, September 2009, p. 9296-9303, Vol. 83, No. 18
0022-538X/09/$08.00+0     doi:10.1128/JVI.00781-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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