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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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Transposons01:24

Transposons

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Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
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DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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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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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis
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Bacteriophage genome-wide transposon mutagenesis.

Alex Chan1, Wearn-Xin Yee1, Deepto Mozumdar1

  • 1Department of Microbiology & Immunology, University of California, San Francisco.

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|December 8, 2025
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Summary

We developed a phage transposon sequencing (TnSeq) platform to identify essential genes in bacteriophages. This scalable technology efficiently maps gene functions for large lytic phages like ΦKZ.

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Area of Science:

  • Bacteriophage functional genomics
  • Molecular biology
  • Genetics

Background:

  • Bacteriophage genomes are large and gene-rich, with many genes of unknown function.
  • Traditional genetic methods for phages are laborious and not scalable.
  • Unbiased functional genomics approaches are crucial for understanding phage biology.

Purpose of the Study:

  • To develop a scalable transposon sequencing (TnSeq) platform for bacteriophage functional genomics.
  • To identify essential and non-essential genes in the jumbo phage ΦKZ.
  • To enable conditional genetic screens and functional analysis of phage genes.

Main Methods:

  • Developed a phage TnSeq platform using a mariner transposase and an anti-CRISPR selectable marker.
  • Employed CRISPR-Cas13a-based enrichment of transposed phages.
  • Utilized pooled sequencing to identify fitness-conferring and dispensable genes in phage ΦKZ.

Main Results:

  • Identified approximately 110 fitness-conferring genes in phage ΦKZ, including essential genes for nucleus formation, protein trafficking, and virion assembly.
  • Generated knockouts for hundreds of non-essential genes.
  • Discovered conditionally essential genes required under specific conditions (clinical isolates, environmental temperature, or nuclease presence).

Conclusions:

  • Phage TnSeq is a facile and scalable technology for defining essential phage genes.
  • This platform enables comprehensive functional genomics and conditional genetic screens in bacteriophages.
  • Provides a broadly applicable resource for studying large lytic phages.