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Related Concept Videos

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...
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...
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.
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Cis-regulatory Sequences02:02

Cis-regulatory Sequences

Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
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...
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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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Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation (BiFC-PALM)
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Coconversion of flanking sequences with homothallic switching.

C McGill1, B Shafer, J Strathern

  • 1Laboratory of Eukaryotic Gene Expression, National Cancer Institute, Frederick Cancer Research Facility, Maryland 21701.

Cell
|May 5, 1989
PubMed
Summary

Homothallic switching in S. cerevisiae replaces mating type DNA. Variable replacement extent and coconversion gradients were observed, suggesting a DNA heteroduplex intermediate during mating type switching.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Yeast Biology

Background:

  • Homothallic switching in Saccharomyces cerevisiae is a process of mating type gene conversion.
  • The mating type locus (MAT) exchanges DNA with silent loci (HML or HMR).

Purpose of the Study:

  • To investigate the variability in DNA replacement extent during homothallic switching.
  • To analyze the gradient of coconversion across the X region of the MAT locus.
  • To explore the implications of flanking site differences for DNA repair mechanisms.

Main Methods:

  • Utilizing restriction site polymorphisms to differentiate between MAT and donor loci.
  • Analyzing DNA sequence replacement patterns during mating type switching.
  • Examining coconversion events relative to HO gene product-mediated double-strand cleavage.

Main Results:

  • Demonstrated variable extent of MAT DNA replacement during homothallic switching.
  • Identified a gradient of coconversion across the X region.
  • Observed coconversion on both sides of the double-strand break.
  • Noted differing flanking sites in daughter cells, indicative of a heteroduplex intermediate.

Conclusions:

  • The extent of DNA replacement during S. cerevisiae mating type switching is not fixed.
  • A DNA heteroduplex intermediate is likely involved in the coconversion process.
  • Understanding these mechanisms provides insights into gene conversion and DNA repair pathways.