Purification of plant viruses and virus coat proteins by high performance liquid chromatography.
High performance liquid chromatography (HPLC) gel filtration has been successfully applied in the purification of elongated and isometric plant viruses. Two different approaches have been tested. In one approach, semi-purified virus particles were dissociated with lithium chloride and the released coat proteins purified by HPLC gel filtration. The purified coat protein was highly immunogenic and gave rise to very specific antisera reacting with intact virus particles as well as with SDS-denatured coat protein monomers. This method is generally applicable only for elongated viruses since many isometric viruses are not dissociated by the lithium chloride treatment. The second approach consisted in gel filtration of native, undissociated virus particles and could be used both with elongated and isometric viruses. Both methods were fast and simple to perform and removed all or most of the contaminating plant proteins as judged by sodium dodecylsulphate gel electrophoresis followed by silver staining or by immunoblotting with antiserum against healthy plant extracts. With both methods the recovery of virus coat protein was about 30% on average.
Albrechtsen
M
M
Plant Protection Centre, Danish Research Service for Plant and Soil Science, Lyngby, Denmark.
Heide
M
M
D000818:Animals; D002213:Capsid / Q000302:isolation & purification*; D002712:Chlorides; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D008094:Lithium; D018021:Lithium Chloride; D009029:Mosaic Viruses / Q000032:analysis / Q000302:isolation & purification; D010942:Plant Viruses / Q000032:analysis / Q000302:isolation & purification*; D010947:Plants, Toxic; D011817:Rabbits; D014026:Nicotiana / Q000382:microbiology
D002712:Chlorides; D008094:Lithium; D018021:Lithium Chloride