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Slippage synthesis of simple sequence DNA.

C Schlötterer1, D Tautz

  • 1Institut für Genetik und Mikrobiologie, Universität München, FRG.

Nucleic Acids Research
|January 25, 1992
PubMed
Summary

Simple sequence DNA synthesis in vitro demonstrates that slippage, a short-range DNA replication error, can generate repetitive DNA motifs. This mechanism likely explains length variations in simple sequence DNA between individuals.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Simple sequence DNA (SSD) comprises repetitive nucleotide motifs crucial for various genomic functions.
  • The in vivo mechanisms generating SSD length polymorphism remain incompletely understood.

Purpose of the Study:

  • To investigate the in vitro synthesis of simple sequence DNA (SSD) using polymerase slippage.
  • To elucidate the role of slippage in the origin and variation of repetitive DNA sequences in vivo.

Main Methods:

  • In vitro synthesis of repetitive di- and trinucleotide motifs using short primers and DNA polymerase.
  • Analysis of synthesis rates and dependence on DNA fragment length and template structure.

Main Results:

  • All types of repetitious di- and trinucleotide motifs were synthesized in vitro.
  • Synthesis rate is sequence-specific, dependent on slippage rate, and independent of fragment length, indicating a short-range effect.
  • Slippage synthesis occurred on a fixed template, mimicking in vivo chromosome replication.

Conclusions:

  • DNA polymerase slippage is a viable mechanism for generating simple sequence DNA motifs in vitro.
  • Slippage during replication is a likely cause of observed length polymorphism in simple sequence DNA stretches among individuals.

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