| 葡萄卷叶病毒外壳蛋白基因的克隆、序列分析及其在大肠杆菌中的表达 |
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| 引用本文： | 张玉满,李云,腊平. 葡萄卷叶病毒外壳蛋白基因的克隆、序列分析及其在大肠杆菌中的表达[J]. 植物病理学报, 2000, 30(3): 232-236 |
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| 作者姓名： | 张玉满 李云 腊平 |
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| 作者单位： | 1 北京林业大学良种繁育研究中心, 北京 100083;2 中国科学院微生物研究所植物生物技术实验室, 北京 100080 |
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| 基金项目： | 国家自然科学基金资助项目 !(39770 6 32 ) |
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| 摘 要： | 从我国感染GLRaV的葡萄韧皮部组织中提取到约18kb的GLRaV-dsRNA。以此为模板,采用RT-PCR方法,扩增到约1.0kb的GLRaV CP基因cDNA片段,并将其克隆到pBluescript Ⅱ SK载体。序列分析表明,GLRaV(中国分离物)CP基因与美国GLRaV-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98.09%。通过大肠杆菌表达载体pBV221构建了GLRaV-CP基因的大肠杆菌蛋白表达克隆,SDS-PAGE电泳分析及WesternBlot结果表明,有一条约35kDa的特异表达蛋白带,与GLRaV-3外壳蛋白大小一致。
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| 关 键 词： | 葡萄卷叶病毒 外壳蛋白基因 克隆 序列 表达 |
| 修稿时间： | 1999-04-20 |
CLONING, SEQUENCING AND EXPRESSING OF COAT PROTEIN GENE OF GRAPEVINE LEAF ROLL ASSOCIATED CLOSTEROVIRUS |
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| ZHANG Yu man ,LI Yun ,TIAN Yan ting ,LA Ping ,CAI Wen qi [WTBZ]. CLONING, SEQUENCING AND EXPRESSING OF COAT PROTEIN GENE OF GRAPEVINE LEAF ROLL ASSOCIATED CLOSTEROVIRUS[J]. Acta Phytopathologica Sinica, 2000, 30(3): 232-236 |
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| Authors: | ZHANG Yu man LI Yun TIAN Yan ting LA Ping CAI Wen qi [WTBZ] |
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| Affiliation: | 1 Research Centre of Improved Cultivar Breeding, Beijing Forest University, Beijing 100083;2 Plant Biotechnology Lab., Institute of Microbiology, The Chinese Academy of Sciences, Beijing 100080, China |
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| 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. |
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| Keywords: | GLRaV 3 coat protein gene cloning sequence expression |
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