Generation of influenza transfectants using purified recombinant nucleocapsid protein

Affinity-purified type A influenza virus nucleocapsid protein expressed by a recombinant baculovirus vector was used in in vitro RNA transcription reactions to create RNP complexes containing a synthetic influenza A virus NS gene. When used in transfection assays, the baculovirus-expressed NP was shown to be biologically active allowing the efficient isolation of transfectant viruses containing the artificially-introduced NS gene. The results demonstrate that NP is the only virion protein necessary in the reconstituted RNP complexes used for transfection thus eliminating the need for purified RNP complexes containing active polymerase.