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Completion sequence and cloning of the infectious cDNA of a chb isolate of cucumber green mottle mosaic virus

机译:黄瓜绿斑驳花叶病毒chb分离株感染性cDNA的完整序列与克隆

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Cucumber green mottle mosaic virus (CGMMV) is an important and widespread seed-borne virus that infects Cucurbitaceous plants. It is a member of the genus Tobamovirus in the family Virgaviridae with a monopartite (+) ssRNA genome. Here we report the complete genome sequence, construction and testing of the infectious clones of a chb isolate of CGMMV. Full-length CGMMV cDNA was cloned into the vector pUC19. The linearized vector containing full-length cDNA was used as template for in vitro transcription, and the synthesized capped transcript was highly infectious in Chenopodium amaranticolor and cucumber (Cucumis sativus). Inoculated plants showed symptoms typical of CGMMV infection. The infectivity was confirmed by mechanical transmission to new plants, RT-PCR and western blot. Progeny virus derived from infectious transcripts had the same biological and biochemical properties as wild-type virus. To our knowledge, this is the first detailed report of a biologically active transcript from CGMMV.
机译:黄瓜绿斑驳花叶病毒(CGMMV)是一种重要且广泛传播的种子传播病毒,可感染葫芦科植物。它是烟草花叶病毒科(Virgaviridae)中的Tobamovirus属成员,具有单部分(+)ssRNA基因组。在这里我们报告完整的基因组序列,CGMMV的chb分离株的感染性克隆的构建和测试。将全长CMGMV cDNA克隆到载体pUC19中。含有全长cDNA的线性化载体被用作体外转录的模板,合成的带帽转录本在Chen形藜和黄瓜(Cucumis sativus)中具有很高的感染力。接种的植物显示出典型的CMGMV感染症状。通过向新植物的机械传播,RT-PCR和Western印迹证实了感染性。源自感染性转录本的子代病毒具有与野生型病毒相同的生物学和生化特性。据我们所知,这是来自CGMMV的生物活性转录本的第一份详细报告。

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