A protocol to study bacteriophage adaptation to new hosts

Raquel Luzon-Hidalgo; Valeria A Risso; Asuncion Delgado; Beatriz Ibarra-Molero; Jose M Sanchez-Ruiz

doi:10.1016/j.xpro.2021.100784

A protocol to study bacteriophage adaptation to new hosts

STAR Protoc. 2021 Aug 26;2(3):100784. doi: 10.1016/j.xpro.2021.100784. eCollection 2021 Sep 17.

Authors

Raquel Luzon-Hidalgo¹, Valeria A Risso¹, Asuncion Delgado¹, Beatriz Ibarra-Molero¹, Jose M Sanchez-Ruiz¹

Affiliation

¹ Departamento de Quimica Fisica, Facultad de Ciencias, Unidad de Excelencia de Quimica Aplicada a Biomedicina y Medioambiente (UEQ), Universidad de Granada, 18071 Granada, Spain.

Abstract

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 engineered E. coli 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).

Keywords: Biophysics; Biotechnology and bioengineering; Evolutionary biology; Microbiology; Model organisms; Molecular biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptation, Biological / genetics
Bacteriophage T7 / genetics
Bacteriophages / genetics*
Escherichia coli / genetics
Genome, Viral / genetics*
High-Throughput Nucleotide Sequencing / methods*
Host-Pathogen Interactions / genetics*
Sequence Analysis, DNA
Virology / methods