TY - JOUR
T1 - Introduction of site-specific mutations into the genome of influenza virus
AU - Enami, Masayoshi
AU - Luytjes, Willem
AU - Krystal, Mark
AU - Palese, Peter
PY - 1990
Y1 - 1990
N2 - 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.
AB - 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.
KW - Negative-sense virus
KW - Rescue of synthetic RNA
KW - Viral selection system
UR - http://www.scopus.com/inward/record.url?scp=0025314308&partnerID=8YFLogxK
U2 - 10.1073/pnas.87.10.3802
DO - 10.1073/pnas.87.10.3802
M3 - Article
C2 - 2339122
AN - SCOPUS:0025314308
SN - 0027-8424
VL - 87
SP - 3802
EP - 3805
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 10
ER -