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Related Concept Videos

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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Conservative Site-specific Recombination and Phase Variation02:53

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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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Fixing Double-strand Breaks02:04

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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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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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DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
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Recombineering Homologous Recombination Constructs in Drosophila
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DNA Recombineering Applications.

Nara Figueroa-Bossi1, Roberto Balbontín2, Lionello Bossi3

  • 1Université Paris-Saclay, CEA, CNRS, Institut de Biologie Intégrative de la Cellule (I2BC), 91190 Gif-sur-Yvette, France.

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|February 22, 2023
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Summary
This summary is machine-generated.

The λ red recombineering technique precisely modifies bacterial genomes without cloning. This versatile method enables gene function studies through various genetic modifications like point mutations and deletions.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Bacterial genome manipulation is crucial for understanding gene function and regulation.
  • Traditional methods often require intermediate molecular cloning steps, which can be time-consuming.

Purpose of the Study:

  • To introduce and explain the λ red recombineering technique for bacterial genome modification.
  • To highlight the versatility of λ red recombineering for diverse genetic applications.

Main Methods:

  • Utilizes the λ red recombinase system for direct modification of bacterial chromosomal DNA.
  • Bypasses the need for traditional molecular cloning procedures.
  • Enables precise base-pair level alterations.

Main Results:

  • Demonstrates the construction of various genetic modifications including insertion mutants, point mutants, and seamless deletions.
  • Shows the application of the technique for creating reporter and epitope tag fusions.
  • Highlights the utility for generating chromosomal rearrangements.

Conclusions:

  • λ red recombineering offers a powerful and efficient approach for bacterial genome engineering.
  • The technique's adaptability makes it suitable for a wide range of genetic studies and synthetic biology applications.