Further analysis of nucleic acids in purified scrapie prion preparations by improved return refocusing gel electrophoresis

• Published in: Journal of general virology 1992-04, Vol.73 (4), p.1025-1029
• Main Authors: Kellings, Klaus, Meyer, Norbert, Mirenda, Carol, Prusiner, Stanley B, Riesner, Detlev
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; DNA - analysis; Fundamental and applied biological sciences. Psychology; Genes. Genome; Molecular and cellular biology; Molecular genetics; Nucleic Acids - analysis; Oligonucleotides - analysis; Poly A - analysis; Prions - chemistry; Prions - isolation & purification; RNA, Messenger - analysis; Scrapie - etiology
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-73-4-1025

1 Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Universitätsstrasse 1, D-4000 Düsseldorf 1, Germany and 2 Department of Neurology and 3 Department of Biochemistry and Biophysics, University of California, San Francisco, California 93143, U.S.A. Although increasingly unlikely, the possibility of a scrapie-specific nucleic acid carried by infectious prion particles is still unresolved. Return refocusing gel electrophoresis was developed to detect homogeneous and heterogeneous nucleic acids extracted from highly purified scrapie prion preparations. This method was improved with respect to the size range from 13 to 1100 nucleotides (nt) over which analyses could be performed. The yield of nucleic acid, particularly of small DNA oligonucleotides and polyadenylated RNA, was determined after deproteinization and two-phase extraction. Despite extensive nuclease digestions some small polynucleotides remained. Although a scrapie-specific nucleic acid cannot be excluded, the results further define the possible characteristics of a hypothetical molecule. If homogeneous in size, such a molecule would be