Mr. Castrucci et Y. Kawaoka, REVERSE GENETICS SYSTEM FOR GENERATION OF AN INFLUENZA-A VIRUS MUTANTCONTAINING A DELETION OF THE CARBOXYL-TERMINAL RESIDUE OF M2 PROTEIN, Journal of virology, 69(5), 1995, pp. 2725-2728
We established a reverse genetics system for the M gene of influenza A
virus, using amantadine resistance as a selection criterion. Transfec
tion of an artificial M ribonucleoprotein complex of A/Puerto Rico/8/3
4 (H1N1), a naturally occurring amantadine-resistant virus, and superi
nfection with amantadine-sensitive A/equine/Miami/1/63 (H3N8), followe
d 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 io 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 th
at carboxyl-terminal residues of the M2 protein play an important role
in influenza virus replication. The M-gene-based reverse genetics sys
tem will allow the study of different M-gene mutations to achieve a ba
lance between attenuation and virus replication, thus facilitating the
production of live vaccine strains.