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Related Concept Videos

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...
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...
Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
Gene Conversion02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...

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Related Experiment Video

Updated: Jun 23, 2026

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
07:55

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

Recombination between linear double-stranded DNA substrates in vivo.

Kumaran Narayanan1, Edmund Ui-Hang Sim, Nikolai V Ravin

  • 1Department of Genetics and Genomic Sciences, Box 1498, Mount Sinai School of Medicine, 1425 Madison Avenue, EB 14-02, New York, NY 10029, USA. kumaran.narayanan@mssm.edu

Analytical Biochemistry
|May 21, 2009
PubMed
Summary

Recombineering in Escherichia coli now supports linear DNA recombination. This advance allows precise in vivo modification and assembly of large linear DNA constructs, including viral genomes and artificial chromosomes.

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Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
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Recombineering Homologous Recombination Constructs in Drosophila
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Recombineering Homologous Recombination Constructs in Drosophila

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Last Updated: Jun 23, 2026

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
07:55

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
06:24

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Published on: February 13, 2019

Recombineering Homologous Recombination Constructs in Drosophila
14:23

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

Area of Science:

  • Molecular Biology
  • Genetics
  • Synthetic Biology

Background:

  • Recombineering in Escherichia coli facilitates targeted integration of linear DNA into circular DNA using homologous sequences.
  • Existing recombineering methods primarily focus on linear-to-circular DNA recombination.

Purpose of the Study:

  • To investigate and demonstrate the capability of recombineering for linear-to-linear DNA recombination in vivo.
  • To assess the accuracy and integrity of large linear DNA molecules after linear/linear recombineering.

Main Methods:

  • Utilized Escherichia coli as the host system for recombineering experiments.
  • Employed linear DNA substrates for recombination assays.
  • Analyzed the recombined products for accuracy and structural integrity.

Main Results:

  • Successfully demonstrated in vivo recombination between two linear DNA substrates (linear/linear recombineering) in E. coli.
  • Showed that linear DNA fragments up to 100 kb can be accurately modified.
  • Confirmed that recombined linear DNA remains intact without unintended rearrangements.

Conclusions:

  • Recombineering technology is versatile and extends to linear/linear DNA recombination in vivo.
  • This novel system enables precise manipulation of large linear DNA elements.
  • Potential applications include engineering of prophages, animal viruses, and assembly of artificial chromosome vectors.