H2N2V1, Main, Exploration, bibRecord, 001841

Eight-plasmid rescue system for influenza A virus

Identifieur interne : 001841 ( Main/Exploration ); précédent : 001840; suivant : 001842

Eight-plasmid rescue system for influenza A virus

Auteurs : Erich Hoffmann [États-Unis] ; Nannan Zhou [États-Unis] ; Robert G. Webster [États-Unis]

Source :

International congress series [ 0531-5131 ] ; 2001.

RBID : ISTEX:06F2C274B31B25392BD51B04E2B6D3D75251C62F

English descriptors

Teeft :
- Allantoic fluid, Cdna, Cell culture, Cocultured cells, Different compartments, Elsevier science, Embryonated eggs, First step, Genetics system, Helper virus infection, Hoffmann, Infectious influenza, Influenza, Influenza vaccines, Influenza virus, Influenza viruses, Internal genes, International congress series, Jude research hospital, Mdck cells, Mrna, Plasmid, Plasmids encoding, Polymerase, Promoter, Protein expression, Recombinant, Recombinant virus, Roche diagnostics, Selection system, Terminator sequences, Transcription, Transcription unit, Transcription units, Transfection, Transfection experiments, Transfection system, Vaccinated population, Vaccine, Vaccine production, Vero cells, Viral, Viral cdna, Viral gene segments, Viral proteins, Virus, Virus particles, Virus titer.

Abstract

Abstract: Plasmid-driven synthesis of viral RNA and protein allows the recovery of infectious influenza virus without the need for helper virus infection. Because no selection system is required for this approach, genetic manipulation of all eight viral gene segments without technical limitations is possible. We have developed a system which requires the construction and transfection of only eight plasmids for the recovery of influenza A viruses. In this DNA transfection system, viral cDNA is inserted between the human RNA polymerase I (pol I) promoter and murine terminator sequences. The entire pol I transcription unit is flanked by an RNA polymerase II (pol II) promoter and a poly(A) site. As a first step to evaluate the utility of this plasmid-based system for the production of vaccines, we generated the master strain A/PR/8/34 (H1N1) currently used for the production of inactivated vaccines entirely from cloned cDNAs. The virus yield as determined by HA-assay after passage of the recombinant virus in eggs was as high as the virus yield of the parental wild-type virus. These results prove that the generated recombinant virus has the same growth properties as the parental egg grown virus and indicate that the eight-plasmid transfection method has the potential to improve currently used methods for the production of vaccine viruses.

Url:

https://api.istex.fr/ark:/67375/6H6-QSJF2021-3/fulltext.pdf

DOI: 10.1016/S0531-5131(01)00375-2

Affiliations:

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Le document en format XML

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<term>Embryonated eggs</term>
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<front><div type="abstract" xml:lang="en">Abstract: Plasmid-driven synthesis of viral RNA and protein allows the recovery of infectious influenza virus without the need for helper virus infection. Because no selection system is required for this approach, genetic manipulation of all eight viral gene segments without technical limitations is possible. We have developed a system which requires the construction and transfection of only eight plasmids for the recovery of influenza A viruses. In this DNA transfection system, viral cDNA is inserted between the human RNA polymerase I (pol I) promoter and murine terminator sequences. The entire pol I transcription unit is flanked by an RNA polymerase II (pol II) promoter and a poly(A) site. As a first step to evaluate the utility of this plasmid-based system for the production of vaccines, we generated the master strain A/PR/8/34 (H1N1) currently used for the production of inactivated vaccines entirely from cloned cDNAs. The virus yield as determined by HA-assay after passage of the recombinant virus in eggs was as high as the virus yield of the parental wild-type virus. These results prove that the generated recombinant virus has the same growth properties as the parental egg grown virus and indicate that the eight-plasmid transfection method has the potential to improve currently used methods for the production of vaccine viruses.</div>
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Eight-plasmid rescue system for influenza A virus

Eight-plasmid rescue system for influenza A virus

Source :

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

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