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Related Experiment Videos

Triplex DNA in plasmids and chromosomes.

J S Lee1, L J Latimer, B L Haug

  • 1Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.

Gene
|October 30, 1989
PubMed
Summary
This summary is machine-generated.

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Circular DNA plasmids with specific sequences can form triplex structures. These triplexes, detectable with a specific antibody, are found in eukaryotes but not E. coli, suggesting a role in chromosome structure.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Circular plasmids with pyrimidine-purine tracts are capable of forming DNA triplex structures.
  • These triplexes can form through intermolecular or intramolecular associations.

Purpose of the Study:

  • To investigate the formation and characteristics of DNA triplexes in circular plasmids.
  • To determine the conditions favoring inter- and intramolecular triplex formation.
  • To assess the presence of triplex structures in different cellular environments.

Main Methods:

  • Plasmid manipulation and addition of synthetic DNA strands (poly(dTC)).
  • Agarose-gel electrophoresis to analyze complex formation.
  • Electron microscopy to visualize triplex structures.

Related Experiment Videos

  • Monoclonal antibody (Jel318) for triplex detection and mobility shift assays.
  • Immunoblotting to detect triplexes in cellular extracts.

    • Main Results:

    • Addition of poly(dTC) induced intermolecular triplex formation in plasmid pTC45, visualized as rosette structures.
    • Intramolecular triplex formation was observed at low pH (pH 5) in plasmids pTC45 and pT463-I.
    • Intermolecular triplexes were stable at pH 8, while intramolecular triplexes required acidic conditions.
    • The monoclonal antibody Jel318 specifically recognized and retarded the mobility of triplex-containing complexes.
    • Triplex structures were detected in eukaryotic cell extracts but not in Escherichia coli extracts.

    Conclusions:

    • DNA triplex formation is a demonstrable phenomenon in circular plasmids under specific conditions.
    • The stability and formation of triplexes are pH-dependent.
    • Triplex structures are associated with eukaryotic cellular machinery and may play a role in eukaryotic chromosome organization.