| 张威1,2;张匀华3;李学湛1;高艳玲1;白艳菊1.应用RT-PCR分子检测技术快速检测大蒜普通潜隐病毒[J].植物保护,2008,34(1):133-137. |
| 应用RT-PCR分子检测技术快速检测大蒜普通潜隐病毒 |
| Rapid detection of Garlic common latent virus by reverse transcription-polymerase chain reaction |
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| DOI： |
| 中文关键词: 大蒜普通潜隐病毒 反转录-聚合酶链式反应 病毒检测 |
| 英文关键词:Garlic common latent virus reverse transcription-polymerase chain reaction virus detection |
| 基金项目: |
| 张威1 2;张匀华3;李学湛1;高艳玲1;白艳菊1 |
| 1.黑龙江省农业科学院植物脱毒苗木研究所 哈尔滨150086;3.黑龙江省农业科学院植物保护研究所 哈尔滨150086;2.黑龙江八一农垦大学植物科技学院 大庆163319 |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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. |
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