【摘要】 从我国感染 GL Ra V的葡萄韧皮部组织中提取到约 18kb的 GL Ra V- ds RNA。以此为模板 ,采用 RT- PCR方法 ,扩增到约 1.0 kb的 GL Ra V CP基因 c DNA片段 ,并将其克隆到p Bluescript IISK载体。序列分析表明 ,GL Ra V(中国分离物 ) CP基因与美国 GL Ra V- 3CP基因核苷酸序列同源性达 99.15% ,推导的氨基酸序列同源性为 98.0 9%。通过大肠杆菌表达载体p BV2 2 1构建了 GLRa V- CP基因的大肠杆菌蛋白表达克隆 ,SDS- PAGE电泳分析及 Western-Blot结果表明 ,有一条约 35k Da的特异表达蛋白带 ,与 GL Ra V- 3外壳蛋白大小一致更多还原

【Abstract】 GLRaV dsRNA, about 18 kb, was extracted from the phloem of grapevine infected with GLRaV in China and the cDNA fragment (ca.1.0 kb) was amplified by RT PCR using dsRNA as template. This product was inserted into pBluescript II SK vector. Analysis of nucleotide and deduced amino acid sequence between this isolate and GLRaV 3, an American isolate, showed 99.15% and 98.09% similarity, respectively. Expression of GLRaV coat protein gene in E.coli showed a 35 kDa unique protein as expected. Result of Western Blot analysis also showed that there is close serum relationship between GLRaV CP Chinese isolate and that of American isolate.更多还原