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应用深度测序技术鉴定平菇球形病毒

DOI: 10.13560/j.cnki.biotech.bull.1985.2014.12.014, PP. 86-91

Keywords: 平菇,深度测序,平菇球形病毒,小干扰RNA,重叠群

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Abstract:

旨在鉴定采集于北京顺义的畸形平菇样品中的病毒,提取总RNA,构建小RNA库,对小RNA库进行深度测序。测序结果表明,与平菇球形病毒(Oystermushroomisometricvirus,OMSV)基因组序列(NC_004560.1)匹配的小RNA共有3075条,分布在OMSV基因组的不同位置,形成一些热点区域。经拼接,获得了3条长度大于500个核苷酸的序列。根据其中覆盖OMSV外壳蛋白(Coatprotein,CP)基因的序列(seq22)中相对保守区域,设计引物,利用RT-PCR扩增获得cp基因部分序列。序列比对表明,其与OMSV相应区域序列相似度为91%,证实了OMSV在畸形平菇样品中的侵染。同时,将深度测序结果与现有4种siRNA预测软件的预测结果进行比较,发现测序结果与软件预测结果之间存在差异,讨论了这一差异出现的原因。

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