Methods and compositions for increasing replication capacity of an influenza virus
In certain aspects, the present invention provides methods for increasing the replication capacity of influenza viruses in hens' eggs and/or cell culture, recombinant and/or reassortant influenza viruses with increased replication capacity, and immunogenic and vaccine compositions comprising such recombinant and/or reassortant influenza viruses. In other aspects, the invention further provides nucleic acids encoding influenza genes associated with increased replication capacity, expression vectors comprising the nucleic acids of the invention, methods for making influenza viruses with increased replication capacity, and kits useful for practice of the methods.
1. A method for increasing the replication capacity of an influenza virus strain MDV-A, comprising altering the amino acid at position 95 of the M1 protein of influenza virus strain MDV-A to lysine (K) and not altering the amino acids at positions 143, 144 and 218, wherein the amino acid at position 143 is valine, the amino acid at position 144 is leucine and the amino acid at position 218 is valine, thereby increasing the replication capacity of the influenza virus.
2. The method of claim 1 , wherein the influenza virus grows to a titer of at least 7.5 log 10 PFU/ml in embryonated eggs.