A vaccine comprising an effective amount of a biologically isolated influenza virus, comprising: i) 8 different gene segments, including a PA viral gene segment, a PB1 viral gene segment, a mutant PB2 viral gene segment, a HA viral gene segment , an NA viral gene segment, an NP viral gene segment, an M viral gene segment (M1 and M2), and an NS viral gene segment (NS1 and NS2), ii) 8 different viral gene segments, including a PA viral gene segment , a PB1 viral gene segment, a mutant PB2 viral gene segment, a HA viral gene segment, an NA viral gene segment (NA and NB), an NP viral gene segment, an M (M1 and BM2) viral gene segment and a segment NS (NS1 and NS2) viral gene, or iii) 7 different gene segments, including a PA viral gene segment, a PB1 viral gene segment, a mutant PB2 viral gene segment, an HEF viral gene segment, an NP viral gene segment, a segment M (M1 and CM2) viral gene, and an NS (NS1 and NS2) viral gene segment； where the mutant PB2 viral gene segment includes 5 'and 3' PB2 non-coding and viral coding incorporation sequences flanking a heterologous nucleotide sequence, and does not include contiguous sequences corresponding to sequences encoding a functional PB2, where the heterologous nucleotide sequence is a gene that encodes a heterologous antigenic protein.Una vacuna comprendiendo una cantidad efectiva de un virus de la gripe aislado biológicamente contenido, comprendiendo: i) 8 segmentos génicos distintos, incluyendo un segmento génico viral PA, un segmento génico viral PB1, un segmento génico viral PB2 mutante, un segmento génico viral HA, un segmento génico viral NA, un segmento génico viral NP, un segmento génico viral M (M1 y M2), y un segmento génico viral NS (NS1 y NS2), ii) 8 segmentos génicos virales distintos, incluyendo un segmento génico viral PA, un segmento génico viral PB1, un segmento génico viral PB2 mutante, un segmento génico viral HA, un segmento génico viral NA (NA y NB), un segmento génico viral NP, un segmento génico vira
