In vitro expression in eukaryotic cells of a prion protein gene cloned from scrapie-infected mouse brain.
It has been proposed that the causative agent of scrapie represents a class of infectious particle that is devoid of nucleic acid and that an altered form of the endogenous prion protein (PrP) is the agent. However, it has been difficult to exclude the possibility that PrP purified from scrapie tissues might be contaminated with a more conventional viral agent. To obtain PrP uncontaminated by scrapie-infected tissues, PrP cDNA cloned from a scrapie-infected mouse brain was expressed in mouse C127 cells in vitro. mRNA and protein encoded by the cloned PrP gene were identified. The expressed PrP polypeptides appeared to be glycosylated and were released from the cell surface into the medium. Homogenates of the cells expressing the cloned PrP gene were inoculated into susceptible mice but failed to induce clinical signs of scrapie. Thus, either PrP is not the transmissible agent of scrapie or the expressed PrP requires additional modification to be infectious.
D000818:Animals; D001923:Brain Chemistry; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D019286:Endopeptidase K; D051379:Mice; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011061:Poly A / Q000235:genetics; D018622:PrP 27-30 Protein; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012608:Scrapie / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D018622:PrP 27-30 Protein; D011061:Poly A; D004247:DNA; D012697:Serine Endopeptidases; D019286:Endopeptidase K