Departamento de Quimica Fisica, Facultad de Ciencias, Unidad de Excelencia de Quimica Aplicada a Biomedicina y Medioambiente (UEQ), Universidad de Granada, 18071 Granada, Spain.
STAR Protoc. 2021 Aug 26;2(3):100784. doi: 10.1016/j.xpro.2021.100784. eCollection 2021 Sep 17.
A general protocol for the experimental assessment of bacteriophage adaptation to new hosts is described. We use as a model system the lytic phage T7 and an strain modified to hamper the recruitment of a known proviral factor. Our protocol includes steps of phage amplification, plaque and liquid lysis assays, and DNA extraction for next-generation sequencing of the viral genome over several rounds of laboratory evolution thus allowing the investigation of the sequence determinants of viral adaptation. For complete information on the generation and use of this protocol, please refer to Luzon-Hidalgo et al. (2021).
描述了一种用于评估噬菌体适应新宿主的实验的通用方案。我们使用裂解噬菌体 T7 和一种经过修饰的 菌株作为模型系统,该菌株被修改以阻止已知前病毒因子的招募。我们的方案包括噬菌体扩增、噬菌斑和液体裂解测定以及用于下一代测序的 DNA 提取等步骤,对病毒基因组进行了几轮实验室进化,从而可以研究病毒适应的序列决定因素。有关该方案的生成和使用的完整信息,请参阅 Luzon-Hidalgo 等人(2021 年)。