PMID: 7535862May 1, 1995Paper

Reverse genetics system for generation of an influenza A virus mutant containing a deletion of the carboxyl-terminal residue of M2 protein

Journal of Virology
M R Castrucci, Y Kawaoka

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.

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Citations

May 2, 2007·Annals of the New York Academy of Sciences·Timothy K W Cheung, Leo L M Poon
Feb 9, 2012·Expert Review of Vaccines·Xavier Saelens
Apr 15, 2008·The Journal of Biological Chemistry·Chunlong MaLawrence H Pinto
Aug 4, 1999·Proceedings of the National Academy of Sciences of the United States of America·G NeumannY Kawaoka
Dec 24, 1996·Proceedings of the National Academy of Sciences of the United States of America·A Bridgen, R M Elliott
Oct 22, 2002·The Journal of General Virology·Gabriele NeumannYoshihiro Kawaoka
May 16, 2006·Journal of Virology·Kiyoko Iwatsuki-HorimotoYoshihiro Kawaoka
Mar 14, 1998·Journal of Virology·E K ParkY Kawaoka
Jan 23, 2020·Cold Spring Harbor Perspectives in Medicine·Gabriele Neumann
Sep 5, 2001·Virology·G Neumann, Y Kawaoka
Jun 1, 1996·Journal of Virology·S PleschkaA García-Sastre

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