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Related Concept Videos

Genomic DNA in Eukaryotes00:58

Genomic DNA in Eukaryotes

Eukaryotes have large genomes compared to prokaryotes. To fit their genomes into a cell, eukaryotic DNA is packaged extraordinarily tightly inside the nucleus. To achieve this, DNA is tightly wound around proteins called histones, which are packaged into nucleosomes that are joined by linker DNA and coil into chromatin fibers. Additional fibrous proteins further compact the chromatin, which is recognizable as chromosomes during certain phases of cell division.
DNA Packaging00:58

DNA Packaging

Overview

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Related Experiment Video

Updated: May 25, 2026

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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Assembling large DNA segments in yeast.

Héloïse Muller1, Narayana Annaluru, Joy Wu Schwerzmann

  • 1Department of Environmental Health Sciences, Johns Hopkins University School of Public Health, Baltimore, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 14, 2012
PubMed
Summary
This summary is machine-generated.

Researchers successfully assembled a 40-kb DNA fragment in yeast using homologous recombination. This method builds upon the uracil-specific excision reaction (USER) fusion technique for large-scale synthetic genome construction.

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Area of Science:

  • Synthetic biology
  • Molecular biology
  • Genomics

Background:

  • The uracil-specific excision reaction (USER) fusion method enables assembly of small DNA fragments into larger segments.
  • Scaling up DNA assembly is crucial for large-scale synthetic genome projects like the Sc2.0 yeast genome initiative.
  • Existing methods require efficient in vivo assembly of kilobase-sized DNA fragments into megabase-sized chromosomes.

Purpose of the Study:

  • To describe a method for assembling 3-kb DNA fragments into larger, chromosome-sized DNA pieces.
  • To demonstrate the feasibility of in vivo assembly of large DNA constructs in yeast.
  • To advance the construction of synthetic genomes by enabling the assembly of large DNA segments.

Main Methods:

  • Utilized homologous recombination in Saccharomyces cerevisiae for in vivo DNA assembly.
  • Employed a lithium acetate (LiOAc) transformation protocol.
  • Assembled 3-kb DNA fragments, each with a 750-base pair overlap, into a 40-kb DNA construct.

Main Results:

  • Successfully assembled a 40-kb DNA fragment in yeast.
  • Demonstrated the efficacy of homologous recombination for large DNA fragment assembly in vivo.
  • Validated the use of overlapping DNA fragments for constructing larger genomic structures.

Conclusions:

  • Homologous recombination in yeast is an effective strategy for assembling large DNA fragments.
  • This method is a key step towards the assembly of entire synthetic yeast chromosomes.
  • The described technique facilitates the construction of complex, large-scale synthetic genomes.