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Single-stranded poly(deoxyguanylic acid) associates into double- and triple-stranded structures.

A Dugaiczyk, D L Robberson, A Ullrich

|December 9, 1980

Summary

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Circular plasmid DNA (pBR322) modified with oligo(dG) tails forms stable polymers. These structures, resistant to certain enzymes, can be reversed, regenerating the original DNA.

Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

Circular plasmid DNA (pBR322) is a common tool in molecular biology.
Restriction endonucleases like PstI are used to linearize DNA.
Terminal deoxynucleotidyl transferase (TdT) can add nucleotides to DNA termini.

Purpose of the Study:

To investigate the self-association of oligo(dG) tails added to linearized plasmid DNA.
To characterize the resulting polymeric structures and their properties.
To explore the reversibility of these self-associated structures.

Main Methods:

Linearization of pBR322 plasmid DNA using PstI restriction endonuclease.
Enzymatic extension of 3' termini with oligo(dG) using terminal deoxynucleotidyl transferase (TdT).

Related Experiment Videos

Analysis of DNA structures using electron microscopy and agarose gel electrophoresis.

Main Results:

Oligo(dG) tails of approximately 15 nucleotides (dG15) conferred resistance to single-strand specific endonuclease S1.
Self-association of oligo(dG) tails led to the formation of stable linear, circular, and branched plasmid DNA polymers.
These polymeric structures could be melted at elevated temperatures or in formamide, reverting to the original linear monomeric DNA.

Conclusions:

Oligo(dG) tail length influences DNA structural transitions and enzymatic activity.
Self-association of oligo(dG) tails provides a mechanism for creating stable, higher-order DNA structures.
The observed self-association and subsequent melting demonstrate a controllable method for DNA polymerization and depolymerization.