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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...
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Viral Recombination00:57

Viral Recombination

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.

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Related Experiment Video

Updated: Jun 6, 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

Targeting vector construction through recombineering.

Liviu A Malureanu1

  • 1Department of Pediatrics and Adolescent Medicine, Mayo Clinic College of Medicine, Rochester, MN, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 17, 2010
PubMed
Summary

Recombineering offers a faster and more cost-effective method for generating gene targeting vectors in mouse embryonic stem cells. This technique streamlines the creation of mouse models for studying gene function and human diseases.

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Recombineering Homologous Recombination Constructs in Drosophila
14:23

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

Related Experiment Videos

Last Updated: Jun 6, 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

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
  • Genomics

Background:

  • Gene targeting in mouse embryonic stem cells is crucial for studying gene function and modeling human diseases.
  • Conventional cloning methods for generating targeting vectors are expensive, time-consuming, and limited by factors like restriction enzyme availability and repetitive DNA amplification.
  • Existing methods face challenges in manipulating large DNA fragments and specific genomic regions.

Purpose of the Study:

  • To present a detailed protocol for generating gene targeting vectors using recombineering.
  • To offer a more efficient and accessible alternative to conventional cloning-based methods.
  • To facilitate the creation of genetically modified mice for research.

Main Methods:

  • Utilizes recombineering, a technique based on the homologous recombination function of lambda phage in Escherichia coli.
  • Leverages the efficiency of bacteriophage-based recombination for manipulating DNA sequences as short as 30-50 bases.
  • Employs mouse genomic bacterial artificial chromosome (BAC) libraries for retrieving genomic DNA sequences.

Main Results:

  • Demonstrates a successfully applied protocol for generating targeting vectors via recombineering.
  • Highlights the ability of recombineering to perform insertions, deletions, or mutations in virtually any genomic region.
  • Facilitates the retrieval of genomic DNA from BAC libraries.

Conclusions:

  • Recombineering provides a powerful and versatile tool for gene targeting vector construction.
  • This method overcomes limitations associated with traditional cloning techniques.
  • The described protocol offers a detailed guide for researchers aiming to generate targeting vectors efficiently.