Cell-surface expression of influenza virus haemagglutinin in insect cells using a baculovirus vector. 1986

R D Possee

A baculovirus transfer vector was constructed using a cloned copy of the Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin gene. The ATG translation initiation codon and 32 bases of the polyhedrin mRNA 5' leader sequences were deleted and replaced with a cDNA clone of the A/PR/8/34 influenza virus haemagglutinin (HA) gene. The hybrid HA/polyhedrin gene was transferred to AcNPV and polyhedrin-negative recombinants identified which expressed HA activity. The insect cell-derived HA bound a monoclonal antibody specific for the HA gene in immunofluorescence tests and was shown to be plasma membrane-bound by haemadsorption assays. A 65 kDa protein was identified in infected cells by immunoprecipitation. The mRNA from recombinant-infected cells was analysed and revealed a major transcript of 2.95 kilobases consistent with transcription initiation and termination at the authentic polyhedrin sites. Other transcripts of different length were also detected so it was not possible to conclude which was responsible for producing the protein. The results demonstrated that AcNPV can be used to express plasma membrane-bound virus glycoproteins.

UI MeSH Term Description Entries
D007304 Insect Viruses Viruses infecting insects, the largest family being BACULOVIRIDAE. Insect Virus,Virus, Insect,Viruses, Insect
D007915 Lepidoptera A large order of insects comprising the butterflies and moths.
D009980 Influenza A virus The type species of the genus ALPHAINFLUENZAVIRUS that causes influenza and other diseases in humans and animals. Antigenic variation occurs frequently between strains, allowing classification into subtypes and variants. Transmission is usually by aerosol (human and most non-aquatic hosts) or waterborne (ducks). Infected birds shed the virus in their saliva, nasal secretions, and feces. Alphainfluenzavirus influenzae,Avian Orthomyxovirus Type A,FLUAV,Fowl Plague Virus,Human Influenza A Virus,Influenza Virus Type A,Influenza Viruses Type A,Myxovirus influenzae-A hominis,Myxovirus influenzae-A suis,Myxovirus pestis galli,Orthomyxovirus Type A,Orthomyxovirus Type A, Avian,Orthomyxovirus Type A, Human,Orthomyxovirus Type A, Porcine,Pestis galli Myxovirus,Fowl Plague Viruses,Influenza A viruses,Myxovirus influenzae A hominis,Myxovirus influenzae A suis,Myxovirus, Pestis galli,Myxoviruses, Pestis galli,Pestis galli Myxoviruses,Plague Virus, Fowl,Virus, Fowl Plague
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002462 Cell Membrane The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells. Plasma Membrane,Cytoplasmic Membrane,Cell Membranes,Cytoplasmic Membranes,Membrane, Cell,Membrane, Cytoplasmic,Membrane, Plasma,Membranes, Cell,Membranes, Cytoplasmic,Membranes, Plasma,Plasma Membranes
D005786 Gene Expression Regulation Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation. Gene Action Regulation,Regulation of Gene Expression,Expression Regulation, Gene,Regulation, Gene Action,Regulation, Gene Expression
D005814 Genes, Viral The functional hereditary units of VIRUSES. Viral Genes,Gene, Viral,Viral Gene
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D006389 Hemagglutinins, Viral Specific hemagglutinin subtypes encoded by VIRUSES. Viral Hemagglutinin,Viral Hemagglutinins,Hemagglutinin, Viral
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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