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  

DETECTION OF GRAPEVINE FANLEAF VIRUS AND POTATO VIRUS Y BY cDNA PROBES LABELLED WITH BIOTIN

作　　者：魏梅生[1] 相宁[1] 张作芳[1] 张成良[1] 王晋芳[2] 邱并生[2] 田波[2] 

机构地区：[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

出　　处：《植物病理学报》1995年第4期331-334,共4页Acta Phytopathologica Sinica

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。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.

关 键 词：葡萄扇叶病毒 马铃薯Y病毒 生物素记 病毒 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]