Protocol to isolate temperature-sensitive SARS-CoV-2 mutants and identify associated mutations

Autor: Shinya Okamura, Akiho Yoshida, Paola Miyazato, Mai Matsumoto, Hirotaka Ebina
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: STAR Protocols, Vol 4, Iss 2, Pp 102352- (2023)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2023.102352
Popis: Summary: An inability to proliferate at high temperatures typically gives viruses an attenuated phenotype. Here, we present a protocol to obtain and isolate temperature-sensitive (TS) SARS-CoV-2 strains via 5-fluorouracile-induced mutagenesis. We describe steps for the induction of mutations in the wild-type virus and selection of TS clones. We then show how to identify the mutations associated with the TS phenotype, following forward and reverse genetics strategies.For complete details on the use and execution of this protocol, please refer to Yoshida et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Databáze: Directory of Open Access Journals