Expression of the peplomer glycoprotein of murine coronavirus JHM using a baculovirus vector. 1989

S Yoden, and T Kikuchi, and S G Siddell, and F Taguchi
National Institute of Neuroscience, Tokyo, Japan.

The gene encoding the E2 peplomer glycoprotein of coronavirus mouse hepatitis virus JHM strain (JHMV) has been inserted into the genome of Autographa californica nuclear polyhedrosis baculovirus (AcNPV) in lieu of the coding region of the AcNPV polyhedrin gene. This recombinant virus produced E2 protein in insect cells under the control of the baculovirus polyhedrin promotor. The expressed E2 protein was shown in size and antigenic properties to be similar to the E2 protein produced in mouse cells infected by JHMV. The expressed E2 protein was glycosylated and transported to the cell surface; however, no proteolytic cleavage was detected in insect cells. The sera from rats immunized with partially purified E2 protein derived from insect cells reacted in immunoprecipitation and immunofluorescence experiments with the E2 protein produced in JHMV-infected mouse cells. The antiserum failed to neutralize the infectivity of JHMV. These results suggest that the E2 protein expressed by the recombinant baculovirus in insect cells is similar but not identical to the E2 protein produced in JHMV-infected mouse cells. The inability of the E2 protein expressed in insect cells to produce neutralizing antibody is discussed.

UI MeSH Term Description Entries
D007304 Insect Viruses Viruses infecting insects, the largest family being BACULOVIRIDAE. Insect Virus,Virus, Insect,Viruses, Insect
D009500 Neutralization Tests The measurement of infection-blocking titer of ANTISERA by testing a series of dilutions for a given virus-antiserum interaction end-point, which is generally the dilution at which tissue cultures inoculated with the serum-virus mixtures demonstrate cytopathology (CPE) or the dilution at which 50% of test animals injected with serum-virus mixtures show infectivity (ID50) or die (LD50). Neutralization Test,Test, Neutralization,Tests, Neutralization
D011233 Precipitin Tests Serologic tests in which a positive reaction manifested by visible CHEMICAL PRECIPITATION occurs when a soluble ANTIGEN reacts with its precipitins, i.e., ANTIBODIES that can form a precipitate. Precipitin Test,Test, Precipitin,Tests, Precipitin
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D005455 Fluorescent Antibody Technique Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy. Antinuclear Antibody Test, Fluorescent,Coon's Technique,Fluorescent Antinuclear Antibody Test,Fluorescent Protein Tracing,Immunofluorescence Technique,Coon's Technic,Fluorescent Antibody Technic,Immunofluorescence,Immunofluorescence Technic,Antibody Technic, Fluorescent,Antibody Technics, Fluorescent,Antibody Technique, Fluorescent,Antibody Techniques, Fluorescent,Coon Technic,Coon Technique,Coons Technic,Coons Technique,Fluorescent Antibody Technics,Fluorescent Antibody Techniques,Fluorescent Protein Tracings,Immunofluorescence Technics,Immunofluorescence Techniques,Protein Tracing, Fluorescent,Protein Tracings, Fluorescent,Technic, Coon's,Technic, Fluorescent Antibody,Technic, Immunofluorescence,Technics, Fluorescent Antibody,Technics, Immunofluorescence,Technique, Coon's,Technique, Fluorescent Antibody,Technique, Immunofluorescence,Techniques, Fluorescent Antibody,Techniques, Immunofluorescence,Tracing, Fluorescent Protein,Tracings, Fluorescent Protein
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
D006023 Glycoproteins Conjugated protein-carbohydrate compounds including MUCINS; mucoid, and AMYLOID glycoproteins. C-Glycosylated Proteins,Glycosylated Protein,Glycosylated Proteins,N-Glycosylated Proteins,O-Glycosylated Proteins,Glycoprotein,Neoglycoproteins,Protein, Glycosylated,Proteins, C-Glycosylated,Proteins, Glycosylated,Proteins, N-Glycosylated,Proteins, O-Glycosylated
D006517 Murine hepatitis virus A species of the CORONAVIRUS genus causing hepatitis in mice. Four strains have been identified as MHV 1, MHV 2, MHV 3, and MHV 4 (also known as MHV-JHM, which is neurotropic and causes disseminated encephalomyelitis with demyelination as well as focal liver necrosis). Gastroenteritis Virus, Murine,Hepatitis Virus, Mouse,Mouse Hepatitis Virus,Murine Gastroenteritis Virus,MHV-JHM,Murine coronavirus,Gastroenteritis Viruses, Murine,Hepatitis Viruses, Mouse,Mouse Hepatitis Viruses,Murine Gastroenteritis Viruses,Murine coronaviruses,Murine hepatitis viruses
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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