Cloning and expression of NDV hemagglutinin-neuraminidase cDNA in a baculovirus expression vector system.
The hemagglutinin-neuraminidase (HN) gene of the Hitchner B1 strain of Newcastle disease virus (NDV) was cloned as a cDNA and inserted into a baculovirus expression vector. The recombinant HN (recHN) expressed in Spodoptera frugiperda cells had both hemagglutinating and neuraminidase activities both of which were inhibited by polyclonal anti-NDV sera or a monoclonal antibody (MAb) against HN. Infected insect cells could hemadsorb chicken red blood cells suggesting that the recHN is properly glycosylated and transported to the cell surface. A 67-kDa recHN precursor and a 74-kDa, presumably mature, recHN from infected cells were detected by Western blot analysis and were found to comigrate with similar proteins from NDV-infected chick embryo fibroblast cells. The kinetics of synthesis of recHN was similar to that for polyhedrin and some HN appeared in the extracellular medium. HN was copurified with extracellular virus (ECV) from the extracellular medium and was used to immunize chickens. The anti recHN serum was specific to NDV in both ELISA and Western blot analysis.
Nagy
E
E
Department of Veterinary Microbiology and Immunology, University of Guelph, Ontario, Canada.
Derbyshire
J B
JB
Dobos
P
P
Krell
P J
PJ
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular; D015342:DNA Probes; D004279:DNA, Viral / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D015707:HN Protein / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D006382:Hemadsorption; D006387:Hemagglutination, Viral; D007114:Immunization; D007304:Insect Viruses / Q000235:genetics; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000096:biosynthesis / Q000235:genetics; D009522:Newcastle disease virus / Q000201:enzymology / Q000235:genetics* / Q000276:immunology; D009693:Nucleic Acid Hybridization; D015183:Restriction Mapping
D015342:DNA Probes; D004279:DNA, Viral; D015707:HN Protein; D009439:Neuraminidase