利用细菌人工染色体制备重组SARS-CoV-2

Current Protocols in Microbiology Pub Date : 2020-10-13 DOI:10.1002/cpmc.126

Kevin Chiem, Chengjin Ye, Luis Martinez-Sobrido

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引用次数: 16

摘要

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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Generation of Recombinant SARS-CoV-2 Using a Bacterial Artificial Chromosome

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

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Current Protocols in Microbiology Immunology and Microbiology-Parasitology

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期刊介绍： Current Protocols in Microbiology provides detailed, step-by-step instructions for analyzing bacteria, animal and plant viruses, fungi, protozoans and other microbes. It offers updated coverage of emerging technologies and concepts, such as biofilms, quorum sensing and quantitative PCR, as well as proteomic and genomic methods. It is the first comprehensive source of high-quality microbiology protocols that reflects and incorporates the new mandates and capabilities of this robust and rapidly evolving discipline.

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