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Related Concept Videos

Mechanism of Conjugation01:19

Mechanism of Conjugation

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Bacterial conjugation is a mechanism of horizontal gene transfer that enables the exchange of genetic material between bacterial cells through direct contact. This process is facilitated by a donor cell carrying a conjugative plasmid, which encodes genes necessary for pilus formation, DNA replication, and transfer. The conjugative plasmid plays a central role in initiating and executing the transfer of genetic material.The tra region of the conjugative plasmid encodes proteins responsible for...
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Conjugation is a form of horizontal gene transfer that primarily occurs in bacteria and some archaea, promoting genetic diversity and adaptation. Bacteria can acquire resistance genes through conjugative plasmids, allowing them to survive antibiotic treatments that would otherwise be lethal. This process involves direct contact between cells through specialized structures such as the sex pilus and is mediated by conjugative plasmids, including the F (fertility) factor.Conjugation requires...
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The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...
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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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Related Experiment Video

Updated: Sep 8, 2025

Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation
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Mating pair stabilization mediates bacterial conjugation species specificity.

Wen Wen Low1,2, Joshua L C Wong1,2, Leticia C Beltran3

  • 1MRC Centre for Molecular Microbiology and Infection, Imperial College, London, UK.

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|June 13, 2022
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Summary

Bacterial conjugation uses outer membrane protein TraN variants to bind specific recipient receptors, enabling DNA transfer and plasmid spread. This interaction dictates conjugation specificity and aids in predicting resistance plasmid distribution.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Structural Biology

Background:

  • Bacterial conjugation is a key mechanism for horizontal gene transfer, spreading antibiotic resistance and virulence factors via plasmids.
  • Plasmid-encoded outer membrane (OM) protein TraN facilitates DNA transfer by interacting with recipient cell surface receptors.
  • Understanding TraN-receptor interactions is crucial for controlling the spread of antimicrobial resistance.

Purpose of the Study:

  • To characterize the molecular mechanisms of TraN variants and their cognate outer membrane receptors in bacterial conjugation.
  • To establish a classification system for TraN homologues based on structural features and receptor binding.
  • To correlate TraN-OM receptor pairings with the distribution of resistance plasmids in clinical isolates.

Main Methods:

  • Cryo-electron microscopy (Cryo-EM) to determine the structure of TraN-OmpK36 complex.
  • Bioinformatic analysis and AlphaFold structural predictions to identify novel TraN variants and their potential receptors.
  • Comparative analysis of TraN-receptor interactions across different bacterial plasmids and species.

Main Results:

  • Identified specific TraN-OM receptor pairings: TraNpKpQIL with OmpK36, TraNR100-1/TraNpSLT with OmpW, and TraNF plasmid with OmpA.
  • Cryo-EM revealed TraNpKpQIL inserts a β-hairpin into OmpK36, stabilizing the mating pair.
  • A fourth TraN variant (TraNδ) was predicted to bind OmpF, expanding the known TraN-receptor repertoire.
  • Devised a classification scheme (TraNα, TraNβ, TraNγ, TraNδ) based on structural similarity and receptor specificity.

Conclusions:

  • Specific TraN-OM receptor interactions are critical for mating pair stabilization and conjugation efficiency, influencing species specificity.
  • The classification of TraN homologues provides a framework for understanding conjugation dynamics.
  • These findings have implications for predicting the spread of antibiotic resistance plasmids within clinical pathogens.