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采自江苏省南京市郊区的小麦病毒病样品提纯后 ,经SDS聚丙烯酰胺凝胶电泳 ,结果表明 ,病毒外壳蛋白 (CP)为单一组分 ,相对分子质量为 36 5× 10 3 。经 1 2g·dL-1琼脂糖凝胶电泳检测 ,病毒核酸有两个组分 ,即RNA1和RNA2。根据AnbeSohn发表的小麦梭条花叶病毒RNA 13′端部分核苷梭序列 ,设计一对引物 ,以提纯样品的RNA1为模板 ,经RT PCR扩增出包含CP基因在内的 1 7kb的特异性目的片段 ,据此鉴定所采集的毒原为小麦梭条花叶病毒 (WSSMV)。
The samples of wheat virus disease collected from the suburbs of Nanjing City, Jiangsu Province were purified by SDS polyacrylamide gel electrophoresis. The results showed that the viral coat protein (CP) was a single component with a relative molecular mass of 36 5 × 10 3. After 1 2g · dL-1 agarose gel electrophoresis, the viral nucleic acid has two components, RNA1 and RNA2. A pair of primers was designed according to the nucleotide sequences of the 13 ’end of the RNA of wheat spindle mosaic virus RNA published by AnbeSohn. The RNA1 of the purified sample was used as a template to amplify the 17 kb specificities including the CP gene by RT PCR The purpose of the fragment, thus identifying the virulent collected wheat spindly mosaic virus (WSSMV).