RNA recombination between persisting pestivirus and a vaccine strain: Generation of cytopathogenic virus and induction of lethal disease
RNA recombination between persisting pestivirus and a vaccine strain: Generation of cytopathogenic virus and induction of lethal disease
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DOI：
10.1128/jvi.75.14.6256-6264.2001
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发表时间：
2001-07-01
影响因子：
5.4
通讯作者：
Thiel, HJ
中科院分区：
文献类型：
--
作者：
Becher, P;Orlich, M;Thiel, HJ
对从一例粘膜病(MD)病例中分离到的牛病毒性腹泻病毒(BVDV)分离株(1741)的分子分析表明，除一个非细胞病变(NCP)株(NCP 1741)外，还有5个不同的病毒亚基因组RNA。对于每个亚基因组，发现了一个大的内部缺失以及一个编码部分核糖体蛋白S27a的插入序列，该序列与N端截短的泛素单体融合在一起。令人惊讶的是，这两个细胞插入加上编码部分NS3和NS4B的侧翼病毒序列与之前描述的BVDV疫苗株RIT(P.Becher，M.Orlich和H.-J.Thiel，J.Virol)的序列有99%的相同。72：8697-8704,1998)，而其余的亚基因组来自NCP 1741的基因组。进一步的分析，包括重组伙伴的分子克隆和核苷酸测序，发现同源和非同源RNA重组都有助于病毒亚基因组的产生。有趣的是，对于另一个来自MD独立病例的CP BVDV分离物(CP 4584)，同样检测到RIT衍生序列元件的插入。与CP 1741相比，CP 4584编码NS3以及部分NS4A和NS4B的基因组区域存在重复。用从嵌合的cDNA结构中转录的RNA转染牛细胞，结果表明，RIT衍生的插入和CP 4584特异性复制的病毒序列代表了CP 4584细胞致病的遗传基础。值得注意的是，回收的cp病毒在细胞培养中传代后，出现了非cp BVDV和一些基因组结构与BVDV 1741相似的病毒亚基因组。
Molecular analysis of a cytopathogenic (cp) bovine viral diarrhea virus (BVDV) isolate (1741) obtained from a case of mucosal disease (MD) led to the identification of five different viral subgenomic RNAs in addition to a noncytopathogenic (noncp) strain (NCP 1741). For each of the subgenomes, a large internal deletion was found together with an inserted sequence encoding part of ribosomal protein S27a fused to an N-terminally truncated ubiquitin monomer. Surprisingly, the two cellular insertions together with flanking viral sequences encoding parts of NS3 and NS4B are > 99% identical to the previously described sequence of BVDV vaccine strain RIT (P. Becher, M. Orlich, and H.-J. Thiel, J. Virol. 72:8697-8704, 1998), while the remainder of the subgenomes is derived from the genome of NCP 1741. Further analyses including molecular cloning and nucleotide sequencing of the recombination partners revealed that both homologous and nonhomologous RNA recombination contributed to the generation of the viral subgenomes. Interestingly, for another cp BVDV isolate (CP 4584) from an independent case of MD, again an insertion of a RIT-derived sequence element was detected. In contrast to CP 1741, for CP 4584 a duplication of the genomic region encoding NS3 and parts of NS4A and NS4B was found. Transfection of bovine cells with RNA transcribed from a chimeric cDNA construct showed that the RIT-derived insertion together with the CP 4584-specific duplication of viral sequences represents the genetic basis of cytopathogenicity of CP 4584. Remarkably, passages of the recovered cp virus in cell culture led to emergence of noncp BVDV and a number of viral subgenomes whose genome organization was similar to that in BVDV 1741.