摘要： 根据马铃薯纺锤块茎类病毒(PSTVd)基因序列设计合成引物,以感病组织和健康组织总RNA为模板,经RTPCR法扩增出全长的cDNA片段,结果从感病组织中扩增出与预期的360bp大小的目标片段,而健康组织无此扩增产物;将其克隆到质粒pGEMT载体上,进行全序列分析。结果表明,与国内外的报道相比较,核苷酸同源率高达98%以上。PCR产物克隆作为RTPCR反应的阳性对照,解决了毒源保存和传播的问题,为进一步进行抗PSTVd基因工程研究打下了良好基础。

关键词: 马铃薯纺锤块茎类病毒, 反转录-聚合酶链式反应, 检测, 克隆, 序列分析

Abstract: A pair of primers w ere designed and synthesized based on PSTVd gene. The ex cepted size 360bpwas amplified by RTvaluePCR from the infected samples,w hile no amplif ied products w ere obtained from thehealthy t issue samples. The unique amplif ied product was then cloned into the pGEMvalueT vector and sequenced.Comparison of the civ il and foreign nucleotide sequence show ed that the homology are 98%. Acting as the posivaluet ive control of RTvaluePCR,the clone of PCR product helped resolve the problem of storing the viroid source andprevent ing it from spreading. This paved a path for breeding the t ransgenic potato resistant to PST Vd.

Key words: PSTVd, RT??PCR, Detection, Cloning, Sequence analysis

中图分类号: 

• S432??