Transmissible gastroenteritis coronavirus gene 7 is not essential but influences in vivo virus replication and virulence.
Transmissible gastroenteritis coronavirus gene 7 is not essential but influences in vivo virus replication and virulence.
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DOI：
10.1016/s0042-6822(02)00096-x
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发表时间：
2003-03-30
期刊：
影响因子：
3.7
通讯作者：
Enjuanes L
中科院分区：
文献类型：
--
作者：
Ortego J;Sola I;Almazán F;Ceriani JE;Riquelme C;Balasch M;Plana J;Enjuanes L
传染性胃肠炎冠状病毒(TGEV)包括8个重叠的基因，这些基因是由3‘端嵌套的含有前导序列的mRNAs表达的。为了便于病毒基因组的遗传操作，通过复制转录调节序列(TRSS)并在每个基因的5‘端引入独特的限制性内切酶位点来分离基因，使用具有感染性的cDNA克隆。重组TGEV(RTGEV)在细胞培养中以与野生型病毒相似的效率复制，并稳定地保持了引入基因组的修改。相反，感染仔猪的肺和肠道中的rTGEV复制水平和毒力显著降低。从构建的重组TGEV7基因中克隆到表达缺失的重组TGEV7(Δ7)，表明TGEV7基因是病毒复制的非必需基因。有趣的是，在体内感染TGEV-Δ7后，肺部和肠道中的病毒复制和毒力都进一步降低，这表明TGEV7基因影响病毒的致病机制。
Transmissible gastroenteritis coronavirus (TGEV) contains eight overlapping genes that are expressed from a 3′-coterminal nested set of leader-containing mRNAs. To facilitate the genetic manipulation of the viral genome, genes were separated by duplication of transcription regulating sequences (TRSs) and introduction of unique restriction endonuclease sites at the 5′ end of each gene using an infectious cDNA clone. The recombinant TGEV (rTGEV) replicated in cell culture with similar efficiency to the wild-type virus and stably maintained the modifications introduced into the genome. In contrast, the rTGEV replication level in the lungs and gut of infected piglets and virulence were significantly reduced. rTGEV in which gene 7 expression was abrogated (rTGEV-Δ7) were recovered from cDNA constructs, indicating that TGEV gene 7 was a nonessential gene for virus replication. Interestingly, in vivo infections with rTGEV-Δ7 showed an additional reduction in virus replication in the lung and gut, and in virulence, indicating that TGEV gene 7 influences virus pathogenesis.
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