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Bacteriophages, or phages, are viruses that specifically infect bacteria, utilizing their genetic material to hijack host cellular machinery for replication. DNA bacteriophages employ single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) genomes. These phages exhibit diverse replication strategies and host interactions, influencing their ecological roles and applications in biotechnology and medicine.ssDNA BacteriophagesssDNA phages, with their small genomes, utilize unique strategies to...
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In contrast to the lytic cycle, phages infecting bacteria via the lysogenic cycle do not immediately kill their host cell. Instead, they combine their genome with the host genome, allowing the bacteria to replicate the phage DNA along with the bacterial genome. The incorporated copy of the phage genome is called the prophage. Some prophages can re-activate and enter the lytic cycle. This often occurs in response to a perturbation, such as DNA damage, but can also transpire in the absence of...
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Bacteriophages, also known as phages, are specialized viruses that infect bacteria. A key characteristic of phages is their distinctive “head-tail” morphology. A phage begins the infection process (i.e., lytic cycle) by attaching to the outside of a bacterial cell. Attachment is accomplished via proteins in the phage tail that bind to specific receptor proteins on the outer surface of the bacterium. The tail injects the phage’s DNA genome into the bacterial cytoplasm. In the...
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The lysogenic cycle is a crucial viral replication strategy that allows bacteriophages to persist within host cells without immediately destroying them. This process is primarily observed in temperate phages, such as bacteriophage lambda (λ), which infects Escherichia coli. The cycle allows the viral genome to persist across bacterial generations while keeping host cells viable.Integration of the Viral GenomeUpon infection, bacteriophage lambda attaches to the bacterial surface and injects...
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Bacteriophages, or phages, are viruses that specifically infect bacteria. Among them, T-even bacteriophages, such as T4, exhibit a well-characterized lytic replication cycle in Escherichia coli (E. coli). This process ensures the rapid proliferation of the virus while ultimately leading to the destruction of the bacterial host.Attachment and DNA InjectionThe infection process begins with the recognition and binding of the T4 phage to the E. coli cell surface. Tail fibers of the phage...
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A protocol to study bacteriophage adaptation to new hosts.

Raquel Luzon-Hidalgo1, Valeria A Risso1, Asuncion Delgado1

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

STAR Protocols
|September 6, 2021
PubMed
Summary

This study details a protocol for tracking bacteriophage adaptation to new bacterial hosts. Researchers used laboratory evolution with phage T7 and engineered E. coli to identify genetic changes driving viral adaptation.

Keywords:
BiophysicsBiotechnology and bioengineeringEvolutionary biologyMicrobiologyModel organismsMolecular biology

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Area of Science:

  • Microbiology
  • Virology
  • Molecular Biology

Background:

  • Bacteriophage adaptation is crucial for their evolution and interaction with bacterial hosts.
  • Understanding viral adaptation mechanisms requires robust experimental methods.
  • Previous studies have explored phage-host interactions, but a standardized protocol for adaptation assessment is needed.

Purpose of the Study:

  • To describe a general experimental protocol for assessing bacteriophage adaptation to new hosts.
  • To investigate the sequence determinants of viral adaptation using laboratory evolution.
  • To provide a framework for studying phage adaptation in a controlled laboratory setting.

Main Methods:

  • The protocol utilizes the lytic phage T7 and an engineered Escherichia coli strain.
  • Key steps include phage amplification, plaque and liquid lysis assays, and DNA extraction.
  • Next-generation sequencing is employed to analyze the viral genome after several rounds of laboratory evolution.

Main Results:

  • The described protocol allows for the experimental assessment of bacteriophage adaptation.
  • It facilitates the investigation of genetic changes in the viral genome over time.
  • The method enables the identification of sequence determinants responsible for adaptation to modified bacterial hosts.

Conclusions:

  • This protocol provides a systematic approach to study bacteriophage adaptation to novel hosts.
  • It is a valuable tool for understanding the genetic basis of viral evolution.
  • The methodology can be applied to various phage-host systems to elucidate adaptation mechanisms.