Use of Triton X-100 and Sephacryl S-500 HR for the Purification of Cymbidium Mosaic Virus from Orchid Plants

  • Han, Jung-Heon (Division of Applied Biology and Chemistry, College of Agriculture and Life Sciences, Seoul National University, Suwon) ;
  • La, Yong-Joon (Division of Applied Biology and Chemistry, College of Agriculture and Life Sciences, Seoul National University, Suwon) ;
  • Lee, Cheol-Ho (Department of Biological Engineering, College of Natural Sciences and Technology, Seokkyoung University.)
  • Published : 1999.02.01

Abstract

Cymbidium mosaic virus (CyMV) was purified from CyMV infected orchid plant leaves by Sephacryl S-500 HR column chromatography. Partial purification was done by solubilization with Triton X-100 (alkylphenoxypolyethoxy ethanol) and precipitation with polyethylene glycol (PEG 6,000) followed by ultracentrifugation on 30% sucrose cushion. Based on the spectrophotometric analysis, 33 mg of CyMV could be obtained form 100 g of CyMV-infected orchid plant leaves. The purified CyMV represented one distinct homogeneous band by SDS-PAGE, and electron microscopy revealed that it was highly homogeneous and not fragmented. Bioassay demonstrated that the purified CyMV had a normal infectivity to Chenopodium amaranticolor and orchid plants. Based on these results, the purification method in this work could be served as an improved method for the purification of CyMV and similar viruses with good yield, high purity and native integrity.

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