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{"title":"Generation of Recombinant SARS-CoV-2 Using a Bacterial Artificial Chromosome","authors":"Kevin Chiem, Chengjin Ye, Luis Martinez-Sobrido","doi":"10.1002/cpmc.126","DOIUrl":null,"url":null,"abstract":"<p>SARS-CoV-2, the causative agent of COVID-19, has been responsible for a million deaths worldwide as of September 2020. At the time of this writing, there are no available US FDA−approved therapeutics for the treatment of SARS-CoV-2 infection. Here, we describe a detailed protocol to generate recombinant (r)SARS-CoV-2 using reverse-genetics approaches based on the use of a bacterial artificial chromosome (BAC). This method will allow the production of mutant rSARS-CoV-2—which is necessary for understanding the function of viral proteins, viral pathogenesis and/or transmission, and interactions at the virus-host interface—and attenuated SARS-CoV-2 to facilitate the discovery of effective countermeasures to control the ongoing SARS-CoV-2 pandemic. © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: Generation of recombinant SARS-CoV-2 using a bacterial artificial chromosome</p><p><b>Support Protocol</b>: Validation and characterization of rSARS-CoV-2</p>","PeriodicalId":39967,"journal":{"name":"Current Protocols in Microbiology","volume":"59 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmc.126","citationCount":"16","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.126","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 16
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Abstract
SARS-CoV-2, the causative agent of COVID-19, has been responsible for a million deaths worldwide as of September 2020. At the time of this writing, there are no available US FDA−approved therapeutics for the treatment of SARS-CoV-2 infection. Here, we describe a detailed protocol to generate recombinant (r)SARS-CoV-2 using reverse-genetics approaches based on the use of a bacterial artificial chromosome (BAC). This method will allow the production of mutant rSARS-CoV-2—which is necessary for understanding the function of viral proteins, viral pathogenesis and/or transmission, and interactions at the virus-host interface—and attenuated SARS-CoV-2 to facilitate the discovery of effective countermeasures to control the ongoing SARS-CoV-2 pandemic. © 2020 Wiley Periodicals LLC.
Basic Protocol : Generation of recombinant SARS-CoV-2 using a bacterial artificial chromosome
Support Protocol : Validation and characterization of rSARS-CoV-2
利用细菌人工染色体制备重组SARS-CoV-2
截至2020年9月，COVID-19的病原体SARS-CoV-2已在全球造成100万人死亡。在撰写本文时，还没有获得美国FDA批准的治疗SARS-CoV-2感染的药物。在这里，我们描述了一种使用基于细菌人工染色体(BAC)的反向遗传学方法生成重组(r)SARS-CoV-2的详细方案。这种方法将允许生产突变型SARS-CoV-2(这对于了解病毒蛋白的功能、病毒发病机制和/或传播以及病毒-宿主界面的相互作用是必要的)和减毒型SARS-CoV-2，以促进发现有效的对策来控制正在进行的SARS-CoV-2大流行。©2020 Wiley期刊公司基本方案:利用细菌人工染色体生成重组SARS-CoV-2支持方案:SARS-CoV-2的验证和表征
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