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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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Related Experiment Video

Updated: Jun 26, 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

A new positive/negative selection scheme for precise BAC recombineering.

Shuwen Wang1, Yuanjun Zhao, Melanie Leiby

  • 1Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine Hershey, Hershey, PA 17033, USA.

Molecular Biotechnology
|January 23, 2009
PubMed
Summary

A new dual-selection marker speeds up bacterial artificial chromosome (BAC) recombineering. This method precisely modifies large DNA constructs, offering a faster, cost-effective alternative for genetic engineering and molecular studies.

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Recombineering Homologous Recombination Constructs in Drosophila
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Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

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Last Updated: Jun 26, 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
  • Genetic Engineering
  • Microbiology

Background:

  • Recombineering enables modification of large DNA constructs without restriction enzymes.
  • Bacterial artificial chromosomes (BACs) are crucial for genetic engineering and studying chromosomal elements.

Purpose of the Study:

  • To develop a novel selection scheme for bacterial artificial chromosome (BAC) recombineering.
  • To provide a faster and more cost-effective method for precise DNA modification.

Main Methods:

  • Constructed a dual kanamycin and streptomycin selection marker using kanamycin resistance and bacterial rpsL (+) genes.
  • Employed a two-step procedure involving positive selection (kanamycin resistance) and counter-selection (streptomycin sensitivity) for precise BAC modification.

Main Results:

  • The dual marker system facilitated precise modifications in BAC constructs without residual sequences.
  • The new method demonstrated efficiency comparable to the existing galK method.
  • The developed technique offers a faster and more economical alternative to the galK method.

Conclusions:

  • The dual kanamycin and streptomycin selection marker is an effective tool for BAC recombineering.
  • This novel approach enhances the efficiency and reduces the cost of modifying large DNA constructs.