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Reverse genetics system for generation of an influenza A virus mutant containing a deletion of the carboxyl-terminal residue of M2 protein.
- Source :
-
Journal of virology [J Virol] 1995 May; Vol. 69 (5), pp. 2725-8. - Publication Year :
- 1995
-
Abstract
- We established a reverse genetics system for the M gene of influenza A virus, using amantadine resistance as a selection criterion. Transfection of an artificial M ribonucleoprotein complex of A/Puerto Rico/8/34 (H1N1), a naturally occurring amantadine-resistant virus, and superinfection with amantadine-sensitive A/equine/Miami/1/63 (H3N8), followed by cultivation in the presence of the drug, led to the generation of a transfectant virus with the A/Puerto Rico/8/34 (H1N1) M gene. With this system, we attempted to generate a virus containing a deletion in an M-gene product (M2 protein). Viruses lacking the carboxyl-terminal Glu of M2, but not those lacking 5 or 10 carboxyl-terminal residues, were rescued in the presence of amantadine. These findings indicate that carboxyl-terminal residues of the M2 protein play an important role in influenza virus replication. The M-gene-based reverse genetics system will allow the study of different M-gene mutations to achieve a balance between attenuation and virus replication, thus facilitating the production of live vaccine strains.
- Subjects :
- Amantadine pharmacology
Amino Acid Sequence
Animals
Base Sequence
Cattle
Cell Line
Cloning, Molecular
DNA Primers genetics
DNA, Complementary genetics
DNA, Viral genetics
Dogs
Drug Resistance, Microbial genetics
Genes, Viral
Influenza A virus drug effects
Influenza A virus physiology
Ion Channels physiology
Molecular Sequence Data
Mutagenesis
Transfection
Viral Matrix Proteins physiology
Virus Replication genetics
Virus Replication physiology
Influenza A virus genetics
Ion Channels genetics
Sequence Deletion
Viral Matrix Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0022-538X
- Volume :
- 69
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of virology
- Publication Type :
- Academic Journal
- Accession number :
- 7535862
- Full Text :
- https://doi.org/10.1128/JVI.69.5.2725-2728.1995