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Methods Mol Biol. 2017;1602:251-273. doi: 10.1007/978-1-4939-6964-7_16.
Plasmid-Based Reverse Genetics of Influenza A Virus.
Perez DR1, Angel M2, Gonzalez-Reiche AS2, Santos J3, Obadan A3, Martinez-Sobrido L4.
Author information
Abstract
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.
KEYWORDS:
Bidirectional plasmids; Influenza A virus; Plasmid-based reverse genetics; Recombinant influenza A virus; Virus rescue approaches
PMID: 28508225 DOI: 10.1007/978-1-4939-6964-7_16
Plasmid-Based Reverse Genetics of Influenza A Virus.
Perez DR1, Angel M2, Gonzalez-Reiche AS2, Santos J3, Obadan A3, Martinez-Sobrido L4.
Author information
Abstract
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.
KEYWORDS:
Bidirectional plasmids; Influenza A virus; Plasmid-based reverse genetics; Recombinant influenza A virus; Virus rescue approaches
PMID: 28508225 DOI: 10.1007/978-1-4939-6964-7_16