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Isolation of Fidelity Variants of RNA Viruses and Characterization of Virus Mutation Frequency
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隔离温度敏感的SARS-CoV-2突变体和识别相关突变的协议.

Shinya Okamura1, Akiho Yoshida1, Paola Miyazato1

  • 1Virus Vaccine Group, BIKEN Innovative Vaccine Research Alliance Laboratories, Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Suita, Osaka, Japan; The Research Foundation for Microbial Diseases of Osaka University, Suita, Osaka, Japan.

STAR protocols
|June 10, 2023
PubMed
概括

这项研究介绍了一种方法,使用化学突变产生来创建温度敏感 (TS) 的SARS-CoV-2菌株. 这些TS病毒株对于开发更安全的疫苗候选人和抗病毒研究至关重要.

关键词:
微生物学 微生物学分子生物学分子生物学

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科学领域:

  • 病毒学 病毒学
  • 分子生物学分子生物学
  • 遗传学 是一个遗传学.

背景情况:

  • 病毒衰减通常与无法在更高的温度下复制有关.
  • 开发安全的SARS-CoV-2 (严重急性呼吸系统综合征冠状病毒2) 菌株对于疫苗和治疗研究至关重要.

研究的目的:

  • 为生成和分离温度敏感 (TS) SARS-CoV-2菌株提出详细的协议.
  • 概述识别TS表型的遗传突变的方法.

主要方法:

  • 利用5-甲诱导的突变发生来引入突变到野生型的SARS-CoV-2.
  • 采用选择策略来分离TS病毒克隆.
  • 应用前向和逆向遗传学的方法来精确确定致病突变.

主要成果:

  • 成功诱导导致温度敏感SARS-CoV-2的突变.
  • 单独的TS病毒克隆的分离和表征.
  • 鉴定赋予温度敏感表型的特定突变.

结论:

  • 描述的协议提供了一种可靠的方法来生成TS SARS-CoV-2菌株.
  • 这种方法有助于研究病毒遗传学和开发基于减弱病毒的疫苗或治疗方法.