An improved method for purification of turnip mosaic virus (TuMV) was established. The infected turnip leaves were frozen and homogenized with 0.5M potassium phosphate buffer, pH 7.5, containing 0.01M Na-EDTA and 0.1% thioglycolic acid. The crude sap obtained by low-speed centrifugation was simultaneously added with 1% Triton X-100, 4% polyethylene glycol (MW6, 000) and 0.1M NaCl to precipitate TuMV without tight aggregation of the virus particles. The pellet was resuspended in high molarity (0.5M) potassium phosphate buffer added with 0.01M MgCl2. The resuspension fluid was treated further by one cycle of differential centrifugation foll-owed by sucrose density-gradient centrifugation. The purified TuMV thus obtained showed the following characteristics; O.D.260/O.D.280 ratio of 1.23, max. 260/min. 247 ratio of 1.14 and particle length of 745nm. The yield of purified virus was 1.5-4.0mg/100g infected turnip leaves. Not any contaminants from host materials were observed in the purified preparation under electron microscopy.
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Choi et al. (1977) studied this question.