Amplification, expression, and packaging of foreign gene by influenza virus.
A system is described that allows use of recombinant DNA technology to modify the genome of influenza virus, a negative-strand RNA virus, and to engineer vectors for the expression of foreign genes. Recombinant RNA is expressed from plasmid DNA in which the coding sequence of the influenza A virus NS gene is replaced with that of the chloramphenicol acetyltransferase gene. When transfected with purified influenza A virus polymerase proteins--in the presence of helper virus--the recombinant RNA is amplified, expressed, and packaged into virus particles, which can be passaged several times. The data indicate that the 22 5' terminal and the 26 3' terminal bases of the influenza A virus RNA are sufficient to provide the signals for RNA transcription, RNA replication, packaging of RNA into influenza virus particles.
Luytjes
W
W
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029.
Krystal
M
M
Enami
M
M
Parvin
J D
JD
Palese
P
P
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002642:Chick Embryo; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005784:Gene Amplification; D015870:Gene Expression; D005814:Genes, Viral; D009980:Influenza A virus / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D014162:Transfection
D015345:Oligonucleotide Probes; D012321:DNA-Directed RNA Polymerases