Expression of functional influenza virus A polymerase proteins and template from cloned cDNAS in recombinant vaccinia virus infected cells.
cDNAs containing the coding sequences of influenza type A virus polymerase proteins (PB1, PB2 and PA) and nucleoprotein (NP) have been expressed in mammalian cells by T7 polymerase provided by a recombinant vaccinia virus. The resulting proteins are able to form a complex that can copy a negative sense influenza-like RNA, transcribed from input DNA by the T7 polymerase, into a positive sense RNA that is translated into active chloramphenicol acetyltransferase (CAT). In this system there is no requirement for helper virus or purified viral core proteins, thus it will allow manipulation of all proteins as well as template for studies of replication in influenza virus.
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Department of Microbiology, University of Alabama at Birmingham 35294.
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D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D005822:Genetic Vectors; D009980:Influenza A virus / Q000201:enzymology* / Q000235:genetics; D007668:Kidney; D010957:Plasmids; D014176:Protein Biosynthesis; D012324:RNA-Dependent RNA Polymerase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics*
D018076:DNA, Complementary; D015500:Chloramphenicol O-Acetyltransferase; D012324:RNA-Dependent RNA Polymerase