生物素标记cDNA探针检测苹果锈果类病毒  

Apple Scar Skin Viroids Detection by cDNA Probe Marked with Biotin

作　　者：赵英[1] 牛建新[1] 

机构地区：[1]石河子大学农学院园艺系,新疆石河子832003

出　　处：《西北农业学报》2008年第3期302-307,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金资助项目(30360066);国家科技攻关计划引导项目(2003BA546C);兵团科委项目(NKB02SDXNK01SW)

摘　　要：以含苹果锈果类病毒(ASSVd)全长序列的重组质粒PUC为模板DNA,加入ASSVd类病毒特异性引物,应用聚合酶链式反应技术(PCR)合成了生物素标记的cDNA探针,在斑点杂交反应中,探针最适使用浓度为1/10,探针检测感病的ASSVd类病毒的最大稀释倍数是1/100。利用制备的探针进行田间检测,结果与RT-PCR检测结果一致,阴性对照均无杂交信号出现。Plant samples were collected from Xinjiang in China in order to evaluate the incidence of apple scar skin viroids.Collected leaves and shoots were tested using reverse transcription polymerase chain reaction（RT-PCR） joined by ASSVd differential primer with DNA template of recombining plasmid PUC containing total ASSVd sequence.The results showed that the adequate concentration was 1/10 and the maximum dilution times in the detection of viroids infection was 1/100 in spot hybridizing reaction.The result of field sample detection using the same probe showed that the consistency with TR-PCR was high and there was no expression of negative hybridizing signal.

关 键 词：CDNA探针 检测 苹果锈果类病毒 

分 类 号：S436.611[农业科学—农业昆虫与害虫防治]