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Journal of Virology, November 2005, p. 13811-13816, Vol. 79, No. 21
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.21.13811-13816.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Cloning of the Chicken RNA Polymerase I Promoter and Use for Reverse Genetics of Influenza A Viruses in Avian Cells

Pascale Massin ,^, Pierre Rodrigues,^, Monica Marasescu, Sylvie van der Werf, and Nadia Naffakh^*

Unité de Génétique Moléculaire des Virus Respiratoires, URA 1966 CNRS, Institut Pasteur, Paris, France

Received 25 April 2005/ Accepted 27 July 2005

Reverse genetics techniques to rescue influenza viruses have thus far been based on the use of a human polymerase I (PolI) promoter to direct the synthesis of the eight viral RNAs. They can only be used on cells from primate origin due to the species specificity of the PolI promoter. Here we report the cloning of the chicken PolI promoter sequence and the generation of recombinant influenza virus upon transfection of bidirectional PolI/PolII plasmids in avian cells. Potential contributions of this new reverse genetics system in the fields of influenza virus research and influenza vaccine production are discussed.

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* Corresponding author. Mailing address: Unité de Génétique Moléculaire des Virus Respiratoires, URA CNRS 1966, Institut Pasteur, 25 rue du Dr. Roux, 75724 PARIS Cedex 15, France. Phone: 33-1-45-68-88-11. Fax: 33-1-40-61-32-41. E-mail: nnaffakh{at}pasteur.fr.

P.M. and P.R. contributed equally to this study.

Present address: Unité VIVAC, AFSSA, Site des Croix, BP53, 22440 Ploufragan, France.

Present address: AgileBio-"Team Solutions," 75 Rue de Lourmel, 75015 Paris, France.

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Journal of Virology, November 2005, p. 13811-13816, Vol. 79, No. 21
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.21.13811-13816.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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