Abstract
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, and packaging of RNA into influenza virus particles.
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