NR AHWV
AU Marsh,R.F.; Dees,C.; Castle,B.E.; Wade,W.F.; German,T.L.
TI Purification of the scrapie agent by density gradient centrifugation
QU Journal of General Virology 1984 Feb; 65(2): 415-21
PT journal article
AB Plasma membrane-enriched preparations from scrapie-infected and healthy hamster brains, as well as preparations of neural retina, were sonicated, then separated by rate-zonal sedimentation in 10 to 25% Nycodenz gradients. Gradient fractions were extracted with 0.5% Triton X-100 and re-fractionated by equilibrium density centrifugation in linear 25 to 40% CsCl gradients. Infectivity was highest in a fraction having a density of 1.280 g/ml and which contained a visible band of material. Digestion of the Nycodenz fractions with proteinase K before detergent extraction and CsCl fractionation resulted in a shift in the visible band to a density of 1.235 g/ml with most of the scrapie infectivity remaining at 1.280 g/ml. When labelled with 125I after 40-fold concentration, this 1.280 g/ml CsCl fraction from the proteinase K-treated gradients contained only a single band of protein(s) having a mol. wt. near 30 000. No differences were seen between proteins in healthy or scrapie-infected preparations.
MH Animal; Brain/*microbiology/ultrastructure; Cell Membrane/microbiology/ultrastructure; Centrifugation, Density Gradient/methods; Hamsters; Male; Neurons/*microbiology; Prions/*isolation & purification/ultrastructure; Retina/*microbiology; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.
SP englisch
PO England