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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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.
The recognition sites for Cre recombinase called LoxP...
Coordination of Gene Expression Processes in Bacteria01:29

Coordination of Gene Expression Processes in Bacteria

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...
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
Mismatch Repair01:20

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
Mismatch Repair01:36

Mismatch Repair

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Updated: Jun 18, 2026

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Oligonucleotide recombination in Gram-negative bacteria.

Bryan Swingle1, Eric Markel, Nina Costantino

  • 1United States Department of Agriculture-Agricultural Research Service, Ithaca, NY 14853, USA. Bryan.Swingle@ars.usda.gov

Molecular Microbiology
|December 1, 2009
PubMed
Summary

Synthetic DNA oligonucleotides (oligos) can directly recombine with bacterial chromosomes without needing phage proteins. This discovery advances understanding of DNA recombination and its potential applications in various bacteria.

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Last Updated: Jun 18, 2026

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

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Published on: January 8, 2015

Detection of Horizontal Gene Transfer Mediated by Natural Conjugative Plasmids in E. coli
06:56

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Published on: March 24, 2023

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • Homologous recombination is crucial for DNA repair and genetic diversity.
  • Recombineering, a technique utilizing homologous recombination, enables precise genetic modifications in bacteria.
  • Current recombineering methods often rely on phage-encoded proteins, limiting their application across diverse bacterial species.

Purpose of the Study:

  • To investigate a novel RecA-independent homologous recombination pathway mediated by synthetic DNA oligonucleotides (oligos).
  • To assess the feasibility of using oligo recombination for site-specific integration into bacterial chromosomes.
  • To explore the potential of oligo recombination for developing new recombineering tools applicable to a broad range of bacteria.

Main Methods:

  • Transformation of synthetic single-stranded DNA oligonucleotides into bacterial cells.
  • Testing oligo recombination across four genera of Gram-negative bacteria.
  • Analyzing the influence of oligo concentration, sequence, and length on recombination frequency.

Main Results:

  • Site-specific recombination between synthetic oligos and bacterial chromosomes was achieved without additional phage functions.
  • Oligo recombination was successfully demonstrated in multiple Gram-negative bacterial genera.
  • Oligo concentration and sequence were identified as key factors influencing recombination efficiency, with length being less critical.

Conclusions:

  • Synthetic DNA oligonucleotides can mediate RecA-independent homologous recombination in bacteria.
  • This finding offers a new avenue for developing versatile recombineering strategies applicable to diverse bacterial species.
  • The evolutionary conservation of oligo recombination across archaea, eukaryotes, and bacteria suggests a fundamental biological process.