Search for a scrapie-specific nucleic acid: a progress report.
Scrapie agent contains a proteinaceous component as well as an 'informational' molecule (suggested by the existence of distinct strains of scrapie). These operationally defined entities may be the same molecule, an infectious protein, or distinct, in which case a nucleic acid might encode the genetic information. Purification of scrapie agent enriched a protein, PrPSc, by virtue of its relative protease resistance. There is only a single PrP gene and the primary translation product of PrP mRNA is the same in normal and scrapie-infected brain; therefore the normal PrPC and the protease-resistant isoform, PrPSc, found in scrapie, probably result from different post-translational events. To search for scrapie-specific nucleic acid, globin RNA made in vitro was added to highly purified infectious preparations at a ratio of 10(3) molecules per infectious unit, nucleic acids were isolated and denatured, and cDNA synthesized using random oligonucleotide primers. Clones containing globin-related sequences were identified by in situ hybridization. 150 plaques not hybridizing to the globin probe were isolated. Inserts larger than 50 base pairs were analysed. By hybridization to a globin probe at reduced stringency all but four clones were found to contain small globin related inserts; two of these hybridized to hamster repetitive sequences as shown by Southern blot analysis. The other clones not related to hamster nucleic acids may be derived from unknown sources of contamination or from scrapie-specific nucleic acids.
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D005815:Genetic Code; D008954:Models, Biological; D009696:Nucleic Acids / Q000032:analysis / Q000235:genetics*; D011328:Prions / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology; D014779:Virus Replication
D009696:Nucleic Acids; D011328:Prions; D014764:Viral Proteins