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Feline Calicivirus: Recovery of Wild-Type and Recombinant Viruses after Transfection of cRNA or cDNA Constructs

机译:猫杯状病毒:转染cRNA或cDNA构建体后野生型和重组病毒的恢复

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摘要

The RNA genome of the vaccine strain 2024 of feline calicivirus was cloned as cDNA and analyzed by nucleotide sequencing. A full-length DNA copy of the viral genome was established and proved to be a source of infectious cRNA after in vitro transcription and RNA transfection. Virus could also be recovered when the DNA construct was introduced into cells containing phage T7 RNA polymerase that was provided by vaccinia virus MVA-T7. After insertion of the sequence encoding the green fluorescent protein into the structural protein-encoding region of the infectious cDNA clone, a defective replicon was recovered that was able to replicate autonomously and was packaged into virus particles when the structural proteins were provided in trans.
机译:将猫杯状病毒疫苗株2024的RNA基因组克隆为cDNA,并通过核苷酸测序进行分析。建立了病毒基因组的全长DNA拷贝,并证明是体外转录和RNA转染后传染性cRNA的来源。当将DNA构建体引入含有由牛痘病毒MVA-T7提供的噬菌体T7 RNA聚合酶的细胞时,也可以回收病毒。将编码绿色荧光蛋白的序列插入感染性cDNA克隆的结构蛋白编码区后,回收了一个有缺陷的复制子,该复制子能够自主复制,并且当以反式方式提供结构蛋白时被包装到病毒颗粒中。

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