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Related Concept Videos

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Chromosome Structure

A functional eukaryotic chromosome must contain three elements: a centromere, telomeres, and numerous origins of replication.
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Histone variants are the histone proteins with structural and sequence variations. These variants may be regarded as “mutant” forms that replace their canonical histone counterparts in the nucleosomes. Specific post-translational modifications on the histone variants enable further chromatin complexity and regulate tissue-specific gene expression. The most common histone variants are from histone H2A, H2B, and linker histone H1 families. However, several variants of histone H3 variants are also...

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Related Experiment Video

Updated: May 9, 2026

qPCRTag Analysis - A High Throughput, Real Time PCR Assay for Sc2.0 Genotyping
07:00

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Published on: May 25, 2015

Functional selection and analysis of yeast centromeric DNA.

P Hieter, D Pridmore, J H Hegemann

    Cell
    |October 1, 1985
    PubMed
    Summary

    Researchers isolated yeast DNA fragments that prevent plasmid loss during cell division. This method identified ten centromeres and a key plasmid stability locus, revealing conserved centromere DNA elements.

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    Single-Copy Gene Locus Chromatin Purification in Saccharomyces cerevisiae
    10:33

    Single-Copy Gene Locus Chromatin Purification in Saccharomyces cerevisiae

    Published on: November 17, 2023

    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Genomics

    Background:

    • Autonomously replicating plasmids in yeast often exhibit segregation bias, leading to instability.
    • The SUP11 gene, when overexpressed, is lethal, providing a selectable marker for plasmid stability.
    • Understanding plasmid segregation is crucial for genetic engineering and molecular biology applications.

    Purpose of the Study:

    • To develop a direct selection method for isolating yeast genomic DNA fragments that confer stability to autonomously replicating plasmids.
    • To identify and characterize DNA elements responsible for preventing segregation bias.
    • To analyze the sequence conservation of identified centromeric DNA elements.

    Main Methods:

    • A direct selection strategy utilizing the lethal ochre-suppressing tRNA gene (SUP11) was employed.
    • Yeast genomic DNA fragments were cloned into an autonomously replicating plasmid (ARS) lacking a partition function.
    • Selected DNA fragments were mapped to specific yeast chromosomes using alternating field gel electrophoresis (AFGE) and hybridization.
    • Nucleotide sequence comparison of isolated centromeric DNA fragments was performed.

    Main Results:

    • Eleven distinct yeast genomic DNA fragments were successfully isolated, eliminating segregation bias in ARS plasmids.
    • Ten of these fragments were identified as chromosomal centromeres.
    • One fragment was identified as the cis-acting locus essential for endogenous 2-micron plasmid stability.
    • Sequence analysis revealed conserved DNA elements within the ten isolated centromeres.

    Conclusions:

    • A robust selection method was established for identifying DNA elements that ensure stable plasmid inheritance in yeast.
    • The study identified novel centromeric DNA sequences and a key locus for 2-micron plasmid stability.
    • Conserved sequence features within yeast centromeres were elucidated, providing insights into their structure and function.