A full-length cDNA infectious clone of North American type 1 porcine reproductive and respiratory syndrome virus: Expression of green fluorescent protein in the Nsp2 region
A full-length cDNA infectious clone of North American type 1 porcine reproductive and respiratory syndrome virus: Expression of green fluorescent protein in the Nsp2 region
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DOI：
10.1128/jvi.01032-06
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发表时间：
2006-12-01
影响因子：
5.4
通讯作者：
Nelson, Eric A.
中科院分区：
文献类型：
--
作者：
Fang, Ying;Rowland, Raymond R. R.;Nelson, Eric A.
最近在美国出现的一组独特的北美1型猪繁殖与呼吸综合征病毒（PRRSV）给已经受到北美2型PRRSV严重影响的养猪业带来了新的疾病控制问题。在这项研究中，从低毒力的北美I型PRRSV分离物SD01-08中获得了全长cDNA感染性克隆。体外研究表明，克隆病毒保持了与亲本病毒相似的生长特性。用克隆病毒攻击动物的病毒学、病理学和免疫学观察结果与用亲本病毒和改良活病毒疫苗攻击动物的观察结果相似。为了进一步探索作为表达外源基因的病毒骨干的潜力，将绿色荧光蛋白（GFP）插入到病毒预测Nsp2区348和349个氨基酸位置的独特缺失位点，并通过荧光显微镜观察Nsp2-GFP融合蛋白的表达。该北美1型传染性克隆的获得，为进一步研究美国1型PRRSV的基本病毒生物学和致病机制提供了重要的研究工具。
The recent emergence of a unique group of North American type 1 porcine reproductive and respiratory syndrome virus (PRRSV) in the United States presents new disease control problems for a swine industry that has already been impacted seriously by North American type 2 PRRSV. In this study, a full-length cDNA infectious clone was generated from a low-virulence North American type I PRRSV isolate, SD01-08. In vitro studies demonstrated that the cloned virus maintained growth properties similar to those of the parental virus. Virological, pathological, and immunological observations from animals challenged with cloned viruses were similar to those from animals challenged with the parental virus and a modified live virus vaccine. To further explore the potential use as a viral backbone for expressing foreign genes, the green fluorescent protein (GFP) was inserted into a unique deletion site located at amino acid positions 348 and 349 of the predicted Nsp2 region in the virus, and expression of the Nsp2-GFP fusion protein was visualized by fluorescent microscopy. The availability of this North American type 1 infectious clone provides an important research tool for further study of the basic viral biology and pathogenic mechanisms of this group of type 1 PRRSV in the United States.