Reverse Genetics of Influenza B Viruses

  • Aitor Nogales
  • Daniel R. Perez
  • Jefferson Santos
  • Courtney Finch
  • Luis Martínez-SobridoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1602)


Annual influenza epidemics are caused not only by influenza A viruses but also by influenza B viruses. Initially established for the generation of recombinant influenza A viruses, plasmid-based reverse genetics techniques have allowed researchers the generation of wild type and mutant viruses from full-length cDNA copies of the influenza viral genome. These reverse genetics approaches have allowed researchers to answer important questions on the biology of influenza viruses by genetically engineering infectious recombinant viruses. This has resulted in a better understanding of the molecular biology of influenza viruses, including both viral and host factors required for genome replication and transcription. With the ability to generate recombinant viruses containing specific mutations in the viral genome, these reverse genetics tools have also allowed the identification of viral and host factors involved in influenza pathogenesis, transmissibility, host-range interactions and restrictions, and virulence. Likewise, reverse genetics techniques have been used for the implementation of inactivated or live-attenuated influenza vaccines and the identification of anti-influenza drugs and their mechanism of antiviral activity. In 2002, these reverse genetics approaches allowed also the recovery of recombinant influenza B viruses entirely from plasmid DNA. In this chapter we describe the cloning of influenza B/Brisbane/60/2008 viral RNAs into the ambisense pDP-2002 plasmid and the experimental procedures for the successful generation of recombinant influenza B viruses.

Key words

Influenza B virus Plasmid-based reverse genetics Virus rescue approaches Recombinant influenza B virus Ambisense plasmids 



Influenza B research to L.M.-S. was partially funded by the 2014 University of Rochester Research Award. J.S. and D.R.P. are funded by start-up funds from the University of Georgia and by NIH contract HHSN272201400008C.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Aitor Nogales
    • 1
  • Daniel R. Perez
    • 2
  • Jefferson Santos
    • 2
  • Courtney Finch
    • 3
  • Luis Martínez-Sobrido
    • 1
    Email author
  1. 1.Department of Microbiology and ImmunologyUniversity of Rochester School of Medicine and DentistryRochesterUSA
  2. 2.Department of Population Health, Poultry Diagnostic and Research Center, College of Veterinary MedicineUniversity of GeorgiaAthensUSA
  3. 3.Division of Viral ProductsOffice of Vaccines Research and Review, Center for Biologics Evaluation and Research, US Food and Drug AdministrationSilver SpringUSA

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