Examination of a plasmid-based reverse genetics system for human astrovirus.
Examination of a plasmid-based reverse genetics system for human astrovirus.
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检查基于质粒的人类星状病毒反向遗传学系统。
DOI：
10.1111/1348-0421.12317
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Nakanishi A.
中科院分区：
文献类型：
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作者：
Chapellier B;Tange S;Tasaki H;Yoshida K;Zhou Y;Sakon N;Katayama K;Nakanishi A.
研究了一种基于质粒的人类星状病毒1型（HAstV1）反向遗传系统。在转染到293T细胞后，含有HAstV1表达盒的质粒载体表达了似乎能够复制病毒RNA的星状病毒RNA，正如亚基因组RNA的产生和衣壳蛋白的表达所表明的那样，无论转录RNA的外源5 '端如何。在该系统中制备了可感染Caco - 2细胞的颗粒；然而，它们的传染性远低于显然产生的颗粒数量所预期的传染性。利用Huh‐7细胞作为转染宿主，旨在改善病毒衣壳加工以促进病毒粒子成熟，部分恢复了感染性颗粒形成的效率。我们的研究结果支持这样一种可能性，即DNA转染过程诱导了针对晚期而非早期HAstV1感染阶段的细胞反应。
A plasmid‐based reverse genetics system for human astrovirus type 1 (HAstV1) is examined. Upon transfection into 293T cells, the plasmid vector, which harbors a HAstV1 expression cassette, expressed astroviral RNA that appeared to be capable of viral RNA replication, as indicated by the production of subgenomic RNA and capsid protein expression irrespective of the heterologous 5′ ends of the transcribed RNA. Particles infectious to Caco‐2 cells were made in this system; however, their infectivity was much lower than would be expected from the amount of particles apparently produced. Using Huh‐7 cells as the transfection host with the aim of improving viral capsid processing for virion maturation partially restored the efficiency of infectious particle formation. Our results support the possibility that the DNA transfection process induces a cellular response that targets late, but not early, stages of HAstV1 infection.