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Related Experiment Videos

Cloning maize telomeres by complementation in Saccharomyces cerevisiae

J M Gardiner1, E H Coe, S Chao

  • 1Department of Agronomy, University of Missouri-Columbia 65211, USA.

Genome
|August 1, 1996
PubMed
Summary

Researchers cloned maize telomeric restriction fragments that function as telomeres in yeast. These fragments allowed mapping of chromosome arm endpoints and revealed a potential link between centromeric and telomeric sequences in maize.

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Area of Science:

  • Plant Genetics
  • Molecular Biology
  • Yeast Genetics

Background:

  • Telomeres are essential protective caps at the ends of eukaryotic chromosomes.
  • Understanding telomere structure and function in maize (Zea mays) is crucial for genome stability.
  • Cloning maize telomeric sequences can facilitate studies on chromosome structure and evolution.

Purpose of the Study:

  • To clone and characterize maize telomeric DNA sequences.
  • To investigate the functional activity of cloned maize telomeres in a heterologous system (yeast).
  • To utilize cloned telomeric fragments for mapping chromosome ends and exploring sequence relationships.

Main Methods:

  • Cloning of maize telomeric fragments using yeast artificial chromosome (YAC) transformation.

Related Experiment Videos

  • Hybridization screening with Arabidopsis telomere repeat probes.
  • Bal31 nuclease digestion to confirm telomeric localization.
  • Restriction fragment length polymorphism (RFLP) analysis for chromosome mapping.
  • DNA sequencing of cloned fragments.

    • Main Results:

    • Nine maize telomeric YAC transformants (MTYs) were identified.
    • Cloned fragments (pMTY7SC, pMTY9ER) hybridized to Bal31-sensitive maize DNA fragments, confirming telomeric function.
    • Telomeric RFLPs were detected, enabling the determination of seven chromosome arm endpoints.
    • One clone (pMTY7ER) mapped to centromeric regions of chromosomes 2 and 3.
    • Sequencing revealed CA-rich regions with degenerate telomere repeats in the cloned fragments.

    Conclusions:

    • Maize telomeric DNA fragments were successfully cloned and shown to function as telomeres in yeast.
    • The cloned fragments serve as valuable tools for mapping chromosome termini in maize.
    • A potential association between centromeric and telomeric sequences in maize chromosomes was suggested.