title: Characterization of rabies glycoprotein expressed in yeast.
abstract: The rabies virus surface glycoprotein was synthesized in Saccharomyces cerevisiae using an expression vector which contains an inducible promoter from the copper metallothionein gene. The rabies G protein was also expressed constitutively in yeast when cloned under control of the triose dehydrogenase promoter. Polypeptides of 65-68 kDa, which migrated at the same molecular weight as authentic viral rabies G protein species, were synthesized by yeast transformants as detected by immunoblotting with rabies specific antiserum. In addition, these polypeptides were immunoprecipitated with several rabies G-specific monoclonal antibodies which neutralize virus infectivity. The recombinant rabies G proteins were glycosylated and associated with membranes in yeast. When injected into guinea pigs, yeast extracts containing the rabies G protein protected animals from lethal rabies virus challenge when administered intramuscularly. However, the same material did not protect mice from a lethal rabies intracerebral challenge.
authors.affiliation: Department of Molecular Genetics, SmithKline Beecham Animal Health, King of Prussia, Pennsylvania.
authors.forename: S R
authors.initials: SR
authors.lastname: Klepfer
authors.forename: C
authors.initials: C
authors.lastname: Debouck
authors.forename: J
authors.initials: J
authors.lastname: Uffelman
authors.forename: P
authors.initials: P
authors.lastname: Jacobs
authors.forename: A
authors.initials: A
authors.lastname: Bollen
authors.forename: E V
authors.initials: EV
authors.lastname: Jones
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D006031:Glycosylation / Q000187:drug effects; D006168:Guinea Pigs; D007114:Immunization; D007273:Injections, Intramuscular; D051379:Mice; D010957:Plasmids; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational; D011818:Rabies / Q000517:prevention & control; D011819:Rabies Vaccines / Q000096:biosynthesis / Q000276:immunology; D011820:Rabies virus / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D014170:Transformation, Genetic; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D006023:Glycoproteins; D011819:Rabies Vaccines; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus; D014415:Tunicamycin