title: Immunological characterization of the sheep prion protein expressed as fusion proteins in Escherichia coli.
abstract: The prion protein (PrP) from sheep was produced in large quantities of entire protein in Escherichia coli after fusion with a carboxy-terminal hexahistidine sequence. In contrast, amino-terminal fusion with glutathione S-transferase (GST) revealed a high susceptibility toward cleavage of the protein. Both recombinant proteins were recognised, at variable levels, in Western blots using a panel of antibodies against the 40-56, 89-104, 98-113 and 112-115 sequences of the prion protein, similarly to the abnormal prion protein extracted from scrapie-infected sheep. Interestingly, monoclonal antibody 3F4 was found to react with these three proteins in Western blot.
authors.affiliation: Agence Française de Sécurité Sanitaire des Aliments, Lyon, France. t.baron@lyon.afssa.fr
authors.forename: T G
authors.initials: TG
authors.lastname: Baron
authors.forename: D
authors.initials: D
authors.lastname: Betemps
authors.forename: M H
authors.initials: MH
authors.lastname: Groschup
authors.forename: J Y
authors.initials: JY
authors.lastname: Madec
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006639:Histidine; D011328:Prions / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012756:Sheep
chemical_list: D011328:Prions; D011993:Recombinant Fusion Proteins; D006639:Histidine; D005982:Glutathione Transferase