Papers - SUZUKI Nobuhiro

The use of SDS-polyacrylamide gel electrophoresis to assess purification procedure of three potyviruses infecting cucurbitaceous plants. Reviewed

Suzuki, N, Shirako, Y, Ehara, Y

Annals of the Phytopathological Society of Japan   55 ( 5 )   586 - 593   1989

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：The Phytopathological Society of Japan (PSJ)  

We examined the effects of concentration methods, extraction buffers, clarification reagents, and resuspension buffers on virus yield and purity by using SDS-polyacrylamide gel electrophoresis (SDS-PAGE), so as to establish optimal purification procedures of zucchini yellow mosaic virus (ZYMV), watermelon mosaic virus 2 (WMV2) and the watermelon mosaic virus 1 strain of papaya ringspot virus (PRSV-W). We then investigated the physicochemical properties of the three viruses. SDS-PAGE proved very useful to monitor virus purification in a small scale. The three viruses were purified with similar yields of 5-10 mg per 100 g of infected pumpkin leaves by the respective optimal methods. The procedures basically involved clarification with Triton X-100, differential centrifugation using a 20% sucrose pad, and 20-50% sucrose density gradient centrifugation. From WMV2 and PRSV-W purified preparations, capsid protein (CP) of 34,000 (34K) was predominantly detected in addition to minor degradation products of 27K to 33K, respectively. A breakdown product of 28K was, however, as detectabel as intact CP of 33K from ZYMV purified preparation. The RNAs of WMV2 and PRSV-W, which were a little larger than ZYMV RNA, were indistinguishable in size as evaluated with 1.7% agarose gel electrophoresis in denaturing conditions.

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