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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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.
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Homologous Recombination02:31

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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...
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Gene Conversion02:08

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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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Exon Recombination02:32

Exon Recombination

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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. 
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Related Experiment Video

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Recombineering Homologous Recombination Constructs in Drosophila
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λ Recombination and Recombineering.

Kenan C Murphy1

  • 1Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605.

Ecosal Plus
|May 26, 2016
PubMed
Summary

The bacteriophage lambda (λ) Red system facilitates DNA exchange through homologous recombination. This system is crucial for DNA repair and genetic engineering in bacteria like Escherichia coli.

Area of Science:

  • Molecular Biology
  • Microbial Genetics
  • Bacteriophage Research

Background:

  • The bacteriophage λ Red system has been a model for studying homologous recombination for 50 years.
  • Early studies revealed the necessity of DNA replication and double-stranded DNA ends for Red-promoted recombination in vivo.
  • The λ Red system facilitates the exchange of DNA segments with extended homology.

Purpose of the Study:

  • To review the mechanistic details of the bacteriophage λ Red homologous recombination system.
  • To explain the utilization of the λ Red system in recombineering for genetic modification of Escherichia coli.
  • To highlight the impact of Red system-mediated recombineering on bacterial genomics and metabolic engineering.

Main Methods:

  • Co-infection of genetically marked phages to generate recombinants.

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Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
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  • Expression of λ Red system independently of other λ functions.
  • Utilizing limited homology (∼50 bp) for recombination with the E. coli chromosome.

    • Main Results:

    • Recognition of the requirement for phage DNA replication in Red-promoted recombination.
    • Identification of the critical role of double-stranded DNA ends in Red protein access.
    • Demonstration of the λ Red system's efficiency in promoting recombination with limited homology.

    Conclusions:

    • The bacteriophage λ Red system is a powerful tool for homologous recombination and DNA repair.
    • Recombineering using the λ Red system has revolutionized genetic manipulation in E. coli.
    • Advances in bacterial genomics, metabolic engineering, and eukaryotic genetics are attributed to the efficiency of this system.