| 用生物素标记外壳蛋白基因cDNA探针检测葡萄扇叶及马铃薯Y病毒 |
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| 引用本文： | 魏梅生,王晋芳.用生物素标记外壳蛋白基因cDNA探针检测葡萄扇叶及马铃薯Y病毒[J].植物病理学报,1995,25(4):331-334. |
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| 作者姓名： | 魏梅生 王晋芳 |
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| 作者单位： | 1. 农业部植物检疫实验所, 北京 100029;2. 中国科学院微生物研究所, 北京 100080 |
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| 摘 要： | 以葡萄扇叶病毒(GFV)干u马铃薯Y病毒(PVY)外壳蛋白基因的重组质粒为模板,用聚合酶链式反膨技术(PCR)分别合成了长度为1.5kb和0.75kb的生物素标记双链cDNA探针,在斑点杂交反应中,探针的最适使用浓度为1/100,GFV探针检测GFV-RNA的灵敏度为10pg,检测提纯GFV的灵敏度为25pg,检测感病苋色藜的最大稀释倍数为10000倍;PVY探针检测PVY-RNA的灵敏度为30pg,检测提纯PVY灵敏度为500pg,感病烟草检测的最大稀释度为8000倍,阴性对照均无杂交信号出现。
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| 关 键 词： | 葡萄扇叶病毒 马铃薯Y病毒 生物素标记cDNA探针 斑点杂交 |
DETECTION OF GRAPEVINE FANLEAF VIRUS AND POTATO VIRUS Y BY cDNA PROBES LABELLED WITH BIOTIN |
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| Wei Meisheng Xiang Ning Zhang Zuofang Zhang Chengliang.DETECTION OF GRAPEVINE FANLEAF VIRUS AND POTATO VIRUS Y BY cDNA PROBES LABELLED WITH BIOTIN[J].Acta Phytopathologica Sinica,1995,25(4):331-334. |
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| Authors: | Wei Meisheng Xiang Ning Zhang Zuofang Zhang Chengliang |
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| Institution: | 1. Institute of Plant Quarantine, Ministry of Agriculture, Beijing 100029;2. Institute of Microbiology, Academia Sinica, Beijing 100080 |
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| Abstract: | Both grapevine fanieaf virus and potato virus Y biotin-labelled ds-cDNA probes were synthesized by polymerase chain reaction using recombinant coat protein gene as template. The optimum concentration of ds-cDNA probes were 1 : 100 in dot blot hybridization. The sensitivity of detection for GFV-RNA was 10 pg,partially purified GFV was 25pg,the maximum dilution of GFV-infected Chenopodium amaranticolor leaves extract was 1 : 10000; The sensitivity of detection for PVY-RNA was 30pg, partially purified PVY was 500pg,the maximum dilution of PVY infected Nicotiana tabacum leaves extract was 1 : 8000. There were no any hybridization signales in the negative control. |
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| Keywords: | Grapevine fanleaf virus Potato virus Y Biotin labelled cDNA probes Dot- blot hybridization |
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