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Updated: Aug 10, 2026
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Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
Published on: January 26, 2017
Isolation of the yeast histone octamer
1Department of Cell Biology, Stanford University School of Medicine, CA 94305.
Summary
Researchers developed a new method to extract and purify the yeast histone octamer using a hexahistidine tag. This purified histone octamer is fully active for nucleosome reconstitution, advancing chromatin studies.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Histones are crucial for DNA packaging and gene regulation.
- Efficient purification of the histone octamer is essential for studying chromatin structure and function.
- Existing methods for histone octamer isolation can be complex and yield impure results.
Purpose of the Study:
- To describe a streamlined procedure for extracting and purifying the yeast histone octamer.
- To develop a method that yields a highly pure and functionally active histone octamer.
- To facilitate further research into nucleosome assembly and chromatin dynamics.
Main Methods:
- Employing mechanical disruption or spheroplast formation for cell lysis and nuclear isolation.
- Utilizing a hexahistidine tag inserted into histone H2B for affinity purification.
- High-salt extraction of nuclei and chromatin followed by purification of the tagged histone octamer.
Main Results:
- Successful extraction and purification of the yeast histone octamer to near homogeneity.
- The purified histone octamer, tagged at histone H2B, was obtained efficiently.
- The reconstituted nucleosomes using the purified histone octamer demonstrated full activity.
Conclusions:
- A robust and efficient method for yeast histone octamer purification has been established.
- The hexahistidine-tagged histone octamer is suitable for functional studies, including nucleosome reconstitution.
- This purification technique provides a valuable tool for advancing chromatin biology research.
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