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Nucleic Acids Research, 1981, Vol. 9, No. 16 4043-4059
© 1981

CHEMISTRY

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

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

Institute of Molecular Genetics, USSR Academy of Sciences Kurchatov Sq. 46, Moscow 123182 *Novosibirsk Institute of Organic Chemistry, USSR Academy of Sciences (Siberian Branch) Prospect Nauki 9, Novosibirsk-90, 630090, USSR

+To whom all correspondence should be addressed.

Received July 1, 1981. 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 {lambda} 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.


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