Yount Boyd, Curtis Kristopher M, Fritz Elizabeth A, Hensley Lisa E, Jahrling Peter B, Prentice Erik, Denison Mark R, Geisbert Thomas W, Baric Ralph S
Department of Epidemiology and Microbiology, University of North Carolina, Chapel Hill, NC 27599-7435, USA.
Proc Natl Acad Sci U S A. 2003 Oct 28;100(22):12995-3000. doi: 10.1073/pnas.1735582100. Epub 2003 Oct 20.
A previously undescribed coronavirus (CoV) is the etiologic agent responsible for severe acute respiratory syndrome (SARS). Using a panel of contiguous cDNAs that span the entire genome, we have assembled a full-length cDNA of the SARS-CoV Urbani strain, and have rescued molecularly cloned SARS viruses (infectious clone SARS-CoV) that contained the expected marker mutations inserted into the component clones. Recombinant viruses replicated as efficiently as WT virus and both were inhibited by treatment with the cysteine proteinase inhibitor (2S,3S)-transepoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester. In addition, subgenomic transcripts were initiated from the consensus sequence ACGAAC in both the WT and infectious clone SARS-CoV. Availability of a SARS-CoV full-length cDNA provides a template for manipulation of the viral genome, allowing for the rapid and rational development and testing of candidate vaccines and therapeutics against this important human pathogen.
一种先前未被描述的冠状病毒（CoV）是导致严重急性呼吸综合征（SARS）的病原体。利用一组覆盖整个基因组的连续cDNA，我们组装了SARS-CoV Urbani株的全长cDNA，并拯救了分子克隆的SARS病毒（感染性克隆SARS-CoV），这些病毒包含插入到组成克隆中的预期标记突变。重组病毒的复制效率与野生型病毒一样高，并且两者都受到半胱氨酸蛋白酶抑制剂（2S，3S）-反式环氧琥珀酰-L-亮氨酰胺-3-甲基丁烷乙酯处理的抑制。此外，野生型和感染性克隆SARS-CoV中的亚基因组转录本均从共有序列ACGAAC起始。SARS-CoV全长cDNA的可获得性为操纵病毒基因组提供了模板，从而能够针对这种重要的人类病原体快速合理地开发和测试候选疫苗及治疗方法。
