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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...
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

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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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Related Experiment Video

Updated: May 29, 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

A Gateway MultiSite recombination cloning toolkit.

Lena K Petersen1, R Steven Stowers

  • 1Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana, United States of America.

Plos One
|September 21, 2011
PubMed
Summary

This study introduces modular Gateway MultiSite recombination cloning tools for efficient DNA construct generation. These adaptable entry clones and destination vectors enhance flexibility and versatility in biological experiments across model systems.

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Related Experiment Videos

Last Updated: May 29, 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

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
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Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

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
  • Biotechnology

Background:

  • DNA construct generation is a bottleneck in biological research.
  • Gateway recombination cloning improved upon traditional methods for single DNA fragments.
  • Existing Gateway systems have limitations in flexibility and versatility for multi-fragment cloning.

Purpose of the Study:

  • To develop and present a series of entry clones and a destination vector for multi-fragment Gateway recombination cloning.
  • To enhance flexibility and versatility in DNA construct generation.
  • To provide a model system-independent cloning strategy.

Main Methods:

  • Development of entry clones containing GAL4, QF, UAS, QUAS, eGFP, and mCherry.
  • Design of a destination vector for two, three, and four fragment Gateway MultiSite recombination cloning.
  • Demonstration of in vivo functionality in Drosophila for generating expression clones.

Main Results:

  • Successful generation of modular Gateway MultiSite entry clones and a destination vector.
  • Demonstrated in vivo functionality in Drosophila, creating various expression constructs.
  • Established a starter toolkit of entry clones adaptable to multiple model systems.

Conclusions:

  • The developed Gateway MultiSite cloning strategy offers increased flexibility and versatility.
  • The modular entry clones are model system-independent, facilitating broader application.
  • A comprehensive toolkit and inventory are available for Gateway MultiSite cloning.