A study of the reversibility of helix-coil transition in DNA

M. P. Perelroyzen, V. I. Lyamichev, Yu A. Kalambet, Yu L. Lyubchenko, A. V. Vologodskii

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24 Scopus citations

Abstract

The reversibility of DNA melting has been thoroughly investigated at different ionic strengths. We concentrated on those stages of the process that do not involve a complete separation of the strands of the double helix. The differential melting curves of pBR 322 DNA and a fragment of T7 phage DNA in a buffer containing 0.02M Na+ have been shown to differ substantially from the differential curves of renaturation. Electron-microscopic mapping of pBR 322 DNA at different degrees of unwinding (by a previously elaborated technique) has shown that the irreversibility of melting under real experimental conditions is connected with the stage of forming new helical regions during renaturation. In a buffer containing 0.2M Na+ the melting curves of the DNAs used (pBR 322, a fragment of T7 phage DHA, a fragment of phage λ DNA, a fragment of ØX174 phage DNA) coincide with the renaturation curves, i.e. the process is equilibrium. We have carried out a semi-quantitative analysis of the emergence of irreversibility in the melting of a double helix. The problem of comparing theoretical and experimental melting curves is discussed.

Original languageEnglish (US)
Pages (from-to)4043-4059
Number of pages17
JournalNucleic acids research
Volume9
Issue number16
DOIs
StatePublished - Aug 25 1981

ASJC Scopus subject areas

  • Genetics

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