Purification of equine herpesvirus type 1.
A method for the purification of enveloped infectious equine herpesvirus type 1 (EHV-1) is presented. Virus from cell culture fluids harvested at 48 h post infection was concentrated by sedimentation and partially purified by differential precipitation with ammonium sulphate. The final steps of purification consisted of two cycles of flotation of virus in pre-formed CsCl density gradients. Yields of infectious virus were about 30% (18 to 44%) of that present in starting material. As judged by electron microscopy, mixed radioisotope labelling, and absence of phosphohydrolase, virus preparations possessed a high degree of purity. Sedimentation of EHV-1 into CsCl density gradients resulted in low recovery of infectious virus. Flotation of virus in CsCl gradients, however, was not deleterious to infectivity of viral preparations.
D000645:Ammonium Sulfate; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D006564:Herpesviridae / Q000302:isolation & purification*; D004861:Herpesvirus 1, Equid / Q000201:enzymology / Q000302:isolation & purification* / Q000648:ultrastructure; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism; D014764:Viral Proteins
D014764:Viral Proteins; D011494:Protein Kinases; D010744:Phosphoric Monoester Hydrolases; D000645:Ammonium Sulfate