Introduction of site-specific mutations into the genome of influenza virus.
M Enami, W Luytjes, M Krystal… - Proceedings of the …, 1990 - National Acad Sciences
M Enami, W Luytjes, M Krystal, P Palese
Proceedings of the National Academy of Sciences, 1990•National Acad SciencesWe succeeded in rescuing infectious influenza virus by transfecting cells with RNAs derived
from specific recombinant DNAs. RNA corresponding to the neuraminidase (NA) gene of
influenza A/WSN/33 (WSN) virus was transcribed in vitro from plasmid DNA and, following
the addition of purified influenza virus RNA polymerase complex, was transfected into MDBK
cells. Superinfection with helper virus lacking the WSN NA gene resulted in the release of
virus containing the WSN NA gene. We then introduced five point mutations into the WSN …
from specific recombinant DNAs. RNA corresponding to the neuraminidase (NA) gene of
influenza A/WSN/33 (WSN) virus was transcribed in vitro from plasmid DNA and, following
the addition of purified influenza virus RNA polymerase complex, was transfected into MDBK
cells. Superinfection with helper virus lacking the WSN NA gene resulted in the release of
virus containing the WSN NA gene. We then introduced five point mutations into the WSN …
We succeeded in rescuing infectious influenza virus by transfecting cells with RNAs derived from specific recombinant DNAs. RNA corresponding to the neuraminidase (NA) gene of influenza A/WSN/33 (WSN) virus was transcribed in vitro from plasmid DNA and, following the addition of purified influenza virus RNA polymerase complex, was transfected into MDBK cells. Superinfection with helper virus lacking the WSN NA gene resulted in the release of virus containing the WSN NA gene. We then introduced five point mutations into the WSN NA gene by cassette mutagenesis of the plasmid DNA. Sequence analysis of the rescued virus revealed that the genome contained all five mutations present in the mutated plasmid. The ability to create viruses with site-specific mutations will allow the engineering of influenza viruses with defined biological properties.
National Acad Sciences