Plasmid-Based Reverse Genetics of Influenza A Virus

  • Daniel R. PerezEmail author
  • Matthew Angel
  • Ana Silvia Gonzalez-Reiche
  • Jefferson Santos
  • Adebimpe Obadan
  • Luis Martinez-Sobrido
Part of the Methods in Molecular Biology book series (MIMB, volume 1602)


Influenza A viruses have broad host range with a recognized natural reservoir in wild aquatic birds. From this reservoir, novel strains occasionally emerge with the potential to establish stable lineages in other avian and mammalian species, including humans. Understanding the molecular changes that allow influenza A viruses to change host range is essential to better assess their animal and public health risks. Reverse genetics systems have transformed the ability to manipulate and study negative strand RNA viruses. In the particular case of influenza A viruses, plasmid-based reverse genetics approaches have allowed for a better understanding of, among others, virulence, transmission, mechanisms of antiviral resistance, and the development of alternative vaccines and vaccination strategies. In this chapter we describe the cloning of cDNA copies of viral RNA segments derived from a type A influenza virus into reverse genetics plasmid vectors and the experimental procedures for the successful generation of recombinant influenza A viruses.

Key words

Influenza A virus Plasmid-based reverse genetics Virus rescue approaches Recombinant influenza A virus Bidirectional plasmids 



M.A. is funded by the Intramural Research Program of the NIH, NIAID, A. S. G.-R., J. S., A. O. and D. R. P.’s research is funded by the University of Georgia, NIH contract HHSN272201400008C, and Scientific Cooperative Agreements with ARS-USDA. L.M-S is funded by the University of Rochester and the NH contract HHSN272201400005C.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Daniel R. Perez
    • 3
    Email author
  • Matthew Angel
    • 1
  • Ana Silvia Gonzalez-Reiche
    • 1
  • Jefferson Santos
    • 3
  • Adebimpe Obadan
    • 3
  • Luis Martinez-Sobrido
    • 2
  1. 1.Department of Population Health, Poultry Diagnostic and Research CenterUniversity of GeorgiaAthensUSA
  2. 2.Department of Microbiology and ImmunologyUniversity of Rochester School of Medicine and DentistryRochesterUSA
  3. 3.Department of Population Health, Poultry Diagnostic and Research Center, College of Veterinary MedicineUniversity of GeorgiaAthensUSA

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