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Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
In vitro Mutagenesis01:16

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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 4, 2026

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
07:18

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Published on: May 15, 2018

Marker reconstitution mutagenesis: a simple and efficient reverse genetic approach.

Xie Tang1, Junqi Huang, Anup Padmanabhan

  • 1Temasek Life Sciences Laboratory, National University of Singapore, Republic of Singapore. tangxie@tll.org.sg

Yeast (Chichester, England)
|March 2, 2011
PubMed
Summary

A new method called marker reconstitution mutagenesis creates gene mutation series. This technique uses two markers and homologous recombination for precise genetic modification in yeast and potentially other organisms.

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Last Updated: Jun 4, 2026

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
07:18

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Published on: May 15, 2018

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06:59

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Published on: August 11, 2010

Area of Science:

  • Genetics
  • Molecular Biology
  • Yeast Genetics

Background:

  • Generating diverse mutations in specific genes is crucial for understanding gene function.
  • Existing methods for creating mutational allelic series can be complex or inefficient.

Purpose of the Study:

  • To develop a novel and efficient reverse genetic approach for generating comprehensive mutational allelic series.
  • To establish a method that circumvents limitations of current genetic mutation techniques.

Main Methods:

  • Developed 'marker reconstitution mutagenesis,' a two-step process utilizing two selective markers.
  • Employed homologous recombination to insert a partial marker fragment adjacent to the gene of interest.
  • Introduced precise random mutations into the target gene while reconstituting the second selective marker.

Main Results:

  • Successfully generated mutational allelic series for several genes in the fission yeast, Schizosaccharomyces pombe.
  • Demonstrated the feasibility and efficiency of the marker reconstitution mutagenesis approach.
  • The method allows for precise introduction of mutations and selection of desired alleles.

Conclusions:

  • Marker reconstitution mutagenesis offers a robust and adaptable strategy for creating gene mutation collections.
  • This novel approach is valuable for functional genomics studies in Schizosaccharomyces pombe.
  • The method's adaptability suggests potential applications in a wide range of organisms capable of homologous recombination.