High-efficiency formation of influenza virus transfectants.
cDNA-derived RNAs were introduced into the genomes of influenza viruses by using an improved ribonucleoprotein (RNP) transfection protocol. Up to 10(5) viral transfectants with a novel neuraminidase gene could be obtained by using a 35-mm dish (10(6) cells) for RNP transfection. In addition to genes coding for surface proteins (hemagglutinin and neuraminidase), we also exchanged a gene coding for nonsurface proteins. The cDNA-derived influenza A/PR/8/34 virus NS gene was introduced into a temperature-sensitive mutant with a defect in this gene. We suggest that the term influenza virus transfectant be used for those viruses which are made by RNP transfection with cDNA-derived RNA.
Enami
M
M
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029.
Palese
P
P
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism; D009975:Orthomyxoviridae / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D012261:Ribonucleoproteins / Q000378:metabolism; D014162:Transfection
D004279:DNA, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D012367:RNA, Viral; D012261:Ribonucleoproteins; D009439:Neuraminidase