Surface-active agents for isolation of the core component of avian myeloblastosis virus.
Sixty-one surface-active agents were evaluated in a procedure designed to assess their ability to remove the envelope from the core component of avian myeloblastosis virus (AMV). The procedure consisted of centrifugation of intact AMV through a series of sucrose gradients each containing an upper layer of agent at one of eight concentrations between 0.01 and 10%. The effectiveness of an agent in producing AMV cores was indicated by (i) the appearance of light-scattering bands in the region of core buoyant density in gradient tubes; (ii) the range of surfactant concentration over which these bands appeared; and (iii) an electron microscopy assessment by the negative-staining technique of the relative proportion of core to non-core material in each of these bands. Six nonionic surfactants were selected by this screening method for comparison in regard to recovery of core protein and endogenous ribonucleic acid (RNA)-dependent deoxyribonucleic acid (DNA) polymerase activity, as well as further morphologic evaluation by electron microscopy. The nonionic surfactants of the polyoxyethylene alcohol class (particularly, Sterox SL) were most effective. Nonionic surfactants of the polyoxyethylene alkylphenol class (particularly, Nonidet P-40) were also effective. Sterox SL and Nonidet P-40 each gave a more than fivefold increase in specific activity of endogenous RNA-dependent DNA polymerase, and each gave a low recovery of core protein. Sterox SL did not interfere to the extent that Nonidet P-40 did in procedures which involved spectrophotometric assay at 260 nm. The use of Sterox SL resulted in the least envelope contamination of core preparations by electron microscopy examination, the most recovery of protein and endogenous RNA-dependent DNA polymerase activity, and a core buoyant density in sucrose of 1.27 g/ml.
D001354:Avian Leukosis Virus / Q000032:analysis* / Q000201:enzymology / Q000302:isolation & purification; D002499:Centrifugation, Density Gradient; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D005069:Evaluation Studies as Topic; D008722:Methods; D008854:Microscopy, Electron; D010772:Phosphotungstic Acid; D013053:Spectrophotometry; D013194:Staining and Labeling; D013395:Sucrose; D013501:Surface-Active Agents; D014764:Viral Proteins / Q000302:isolation & purification*
D013501:Surface-Active Agents; D014764:Viral Proteins; D010772:Phosphotungstic Acid; D013395:Sucrose; D004254:DNA Nucleotidyltransferases