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Related Concept Videos

DNA Bacteriophages01:26

DNA Bacteriophages

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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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Lytic Cycle of Bacteriophages01:30

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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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Viral Replication: Lysogenic Cycle01:16

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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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Lysogenic Cycle of Bacteriophages00:43

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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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Viral Replication: Lytic Cycle01:20

Viral Replication: Lytic Cycle

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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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Transduction01:16

Transduction

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Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome...
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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Directed Evolution of Replication-Competent Double-Stranded DNA Bacteriophage toward New Host Specificity.

Jing Liang1, Huibin Zhang2, Yee Ling Tan1

  • 1Strain Engineering, Singapore Institute of Food and Biotechnology Innovation, Singapore 138669, Singapore.

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|January 28, 2022
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Summary

Bacteriophages offer a promising alternative to antibiotics for combating antimicrobial resistance. This study engineered T7 bacteriophage variants with altered host specificity, demonstrating potential for broader therapeutic applications.

Keywords:
directed evolutionphage engineeringphage specificitysynthetic phage platformtail fiber engineering

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Antimicrobial resistance necessitates novel therapeutic strategies.
  • Bacteriophages (phages) show promise as alternatives to antibiotics.
  • Phage therapy requires specific host-bacteriophage matching due to narrow host ranges.

Purpose of the Study:

  • To engineer bacteriophage specificity and improve therapeutic qualities.
  • To develop a method for generating large libraries of phage variants with diverse host specificities.
  • To explore directed evolution for enhancing phage characteristics beyond natural selection.

Main Methods:

  • Utilized directed evolution to create large libraries of replication-competent phage variants from synthetic DNA.
  • Generated a library of over 10^7 T7 bacteriophage tail fiber mutants.
  • Screened mutant libraries for altered host specificity and improved lytic efficiency.

Main Results:

  • Identified T7 phage mutants with broadened host specificity, including lytic activity against *Yersinia enterocolitica*.
  • Discovered mutants exhibiting enhanced lytic efficiency and improved tolerance to lytic conditions.
  • Observed limitations in altering host specificity solely through tail fiber mutagenesis, suggesting other factors are critical.

Conclusions:

  • Directed evolution of bacteriophages is a viable strategy for engineering host specificity and therapeutic traits.
  • Tail fiber modification can broaden phage host range, but other genetic elements may also limit specificity.
  • This approach holds potential for developing tailored phage therapies to combat antimicrobial resistance.