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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

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...
Crossing Over01:30

Crossing Over

Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to allelic diversity in the newly formed daughter cells. In humans, crossing over produces genetically distinct haploid egg and sperm cells that undergo fertilization to produce unique offspring. Before cell division starts, the germ cell’s chromosome(s) undergo duplication in the S phase of the cell cycle. As the cells enter prophase I, duplicated...
Crossing Over01:34

Crossing Over

Unlike mitosis, meiosis aims for genetic diversity in its creation of haploid gametes. Dividing germ cells first begin this process in prophase I, where each chromosome—replicated in S phase—is now composed of two sister chromatids (identical copies) joined centrally.
The homologous pairs of sister chromosomes—one from the maternal and one from the paternal genome—then begin to align alongside each other lengthwise, matching corresponding DNA positions in a process called synapsis.
In order to...
Exon Recombination02:32

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon has three reading...

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Related Experiment Video

Updated: Jul 5, 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

Building larger YACs by recombination.

G A Silverman1

  • 1Children's Hospital and Harvard Medical School, Boston, Massachusetts, USA.

Current Protocols in Human Genetics
|April 23, 2008
PubMed
Summary
This summary is machine-generated.

Reconstruct intact genes from overlapping DNA segments using homologous recombination in yeast. This method utilizes either the meiotic or mitotic phases of the yeast life cycle for efficient gene cloning.

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

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Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
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Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

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Related Experiment Videos

Last Updated: Jul 5, 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

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
08:48

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

Published on: January 26, 2017

Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Genomic Cloning

Background:

  • Large-insert cloning vectors like yeast artificial chromosomes (YACs) often fail to capture entire genomic regions or genes intact.
  • Genomic regions are frequently represented as a collection of overlapping DNA fragments (contigs) rather than a single clone.

Purpose of the Study:

  • To describe protocols for reconstructing intact genes within a single yeast artificial chromosome (YAC) clone.
  • To leverage homologous recombination in Saccharomyces cerevisiae for assembling overlapping DNA segments.

Main Methods:

  • Utilizing homologous recombination in Saccharomyces cerevisiae to splice together overlapping DNA segments.
  • Implementing two distinct protocols: one based on the meiotic phase and another on the mitotic phase of the yeast cell cycle.

Main Results:

  • Successful reconstruction of intact genes from fragmented YAC clones.
  • Demonstration of high-frequency and high-fidelity homologous recombination in yeast for gene assembly.

Conclusions:

  • Homologous recombination in Saccharomyces cerevisiae provides an effective strategy for overcoming limitations of YAC cloning.
  • The described protocols enable the generation of complete gene clones from existing overlapping YAC contigs.