| 马铃薯S病毒外壳蛋白基因的克隆及其在大肠杆菌中的表达 |
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| 引用本文： | 李广存,杨煜,王秀丽,杨元军,毕玉平. 马铃薯S病毒外壳蛋白基因的克隆及其在大肠杆菌中的表达[J]. 园艺学报, 2004, 31(4): 517-519 |
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| 作者姓名： | 李广存 杨煜 王秀丽 杨元军 毕玉平 |
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| 作者单位： | ( 山东省农业科学院生物中心，济南250100； 山东省农业科学院蔬菜研究所，济南250100) |
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| 基金项目： | 农业部“948”项目(201047)，山东省科技攻关项目(99-05) |
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| 摘 要： | 以田间采集的马铃薯病叶中提取的马铃薯病毒s(PVS)总RNA为模板，通过RT—PCR获取长度为890 bp的P 一cp的cDNA，克隆至pGEM—T载体上。酶切回收该基因片段，并构建了该基因的原核表达质粒pBV—pvs。SDS—PAGE凝胶电泳和Western印迹分析表明：P 一印基因在大肠杆菌JM109中可特异地高效表达分子量约33kD的蛋白，且表达蛋白具有良好的抗原活性。利用该表达产物免疫动物家兔，获得的抗血清可用于大田马铃薯s病毒的快速检测。
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| 关 键 词： | 马铃薯 马铃薯s病毒 外壳蛋白基因 RT—PCR 序列分析 Western印迹 抗血清制备 |
| 文章编号： | 0513-353X(2004)04-0517-03 |
Cloning of Potato Virus S Coat Protein Gene and Its Expression in E.coli JM109 |
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| Li Guangcun,Yang Yu,Wang Xiuli,Yang Yuanjun,and Bi Yuping. Cloning of Potato Virus S Coat Protein Gene and Its Expression in E.coli JM109[J]. Acta Horticulturae Sinica, 2004, 31(4): 517-519 |
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| Authors: | Li Guangcun Yang Yu Wang Xiuli Yang Yuanjun and Bi Yuping |
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| Abstract: | Total RNA of potato virus S (PVS) was isolated from diseased potato leaves collected in field, The cDNA encoding PVS coat protein (PVS-cp) was amplified by RT-PCR and cloned into the plas-mid pGEM-T vector. The expression vector of PVS-cp gene was constructed through inserting the cDNA fragment into the expression plasmid pBV220. The results of SDS-PAGE and Western blot analysis showed that the PVS-cp gene highly expressed in E. coli JM109 and the molecular size of the expression product was about 33 kD. The product was used as antigen to immunize rabbits and the specific anti-serum can be used to detect PVS rapidly. |
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| Keywords: | Potato Potato virus S Coat protein gene RT-PCR Sequence analysis Western blot Anti-serum preparation |
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