Purification of barley yellow dwarf virus by gel filtration on Sephacryl S-1000 Superfine.

Barley yellow dwarf virus was purified from infected oats using cellulase to assist virus extraction, clarification of the extract with chloroform-butanol, precipitation of virus by polyethylene glycol and gel filtration of the resulting suspension on Sephacryl S-1000 Superfine. The virus was further purified by sucrose density gradient centrifugation. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and electron microscopy showed that the virus preparation is of sufficient purity for biochemical investigation. A yield of approximately 0.8 mg virus/kg of infected leaf was obtained. The technique is simple and less time-consuming than conventional centrifugation procedures and can be used routinely for purification of a wide range of spherical plant viruses.