摘要
根据大蒜普通潜隐病毒(Garlic common latent virus,GCLV)的外壳蛋白区域的保守序列设计合成一对寡核苷酸引物,以带毒植物的总RNA为模板,进行反转录和PCR扩增,通过反应体系和反应程序的建立与优化,扩增得到长300 bp的目的片段,并将目的片段转入大肠杆菌进行了克隆和序列测定。测序结果与GenBank中其他GCLV相应区域的序列同源性最高达98%。并对其检测的特异性和灵敏度进行了验证,从而建立了快速、灵敏、特异性强的GCLV分子生物学检测方法。
With specific primers designed based on the conserved sequence of Garlic common latent virus （GCLV） coat protein gene, a 0.3-kb fragment was amplified by reverse transcription-polymerase chain reaction （RT-PCR） using the total RNA of the plants infected with GCLV. The fragment was cloned into Escherichia coli GM109 and sequenced. The sequence was compared with the sequences of other GCLV isolates retrieved from GenBank. The results showed that it was highly homologous to that of other isolates, with the highest identity as 98%. The specificity and sensitivity of this detection method were verified. It was a rapid and specific system for GCLV detection.
出处
《植物保护》
CAS
CSCD
北大核心
2008年第1期133-137,共5页
Plant Protection
基金
黑龙江省自然科学基金项目(C2004-05)
关键词
大蒜普通潜隐病毒
反转录-聚合酶链式反应
病毒检测
Garlic common latent virus
reverse transcription-polymerase chain reaction
virus detection
作者简介
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