A baculovirus dual expression vector derived from the Autographa californica nuclear polyhedrosis virus polyhedrin and p10 promoters: co-expression of two influenza virus genes in insect cells.
A baculovirus transfer vector, pAcUW3, was developed to facilitate the insertion of two influenza virus genes, those encoding the haemagglutinin (HA) and neuraminidase (NA) membrane glycoproteins, into the Autographa californica nuclear polyhedrosis virus genome in a single cotransfection experiment. The NA gene was inserted in place of the polyhedrin coding sequences under the control of the polyhedrin promoter, whereas the HA gene was placed under the control of a copy of the p10 promoter at a site upstream of and in opposite orientation to the polyhedrin promoter. After infection of Spodoptera frugiperda cells with the recombinant virus, AcUW3HANA, both HA and NA were expressed in the very late phase of infection and were shown to be functional in appropriate assays. Immunofluorescence assays demonstrated their localization at the surface of infected insect cells. The expression of both foreign genes in the recombinant virus was found to be stable for at least 12 passages in cell culture.
Weyer
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NERC Institute of Virology and Environmental Microbiology, Oxford, U.K.
Possee
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D000818:Animals; D000911:Antibodies, Monoclonal; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D005822:Genetic Vectors; D006386:Hemagglutination Tests; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology; D009980:Influenza A virus / Q000235:genetics*; D008969:Molecular Sequence Data; D009036:Moths / Q000382:microbiology; D009439:Neuraminidase / Q000235:genetics / Q000276:immunology; D000077183:Occlusion Body Matrix Proteins; D010957:Plasmids; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014764:Viral Proteins / Q000235:genetics*; D015678:Viral Structural Proteins
D000911:Antibodies, Monoclonal; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D000077183:Occlusion Body Matrix Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D009439:Neuraminidase