Intramolecular DNA melting between stable helical segments: melting theory and metastable states.

Abstract:

:Melting of DNA in a segment bounded at both ends by regions of greater stability during electrophoresis in denaturing gradient gels show complex properties, not accommodated with standard melting theory. Compact bands of some DNA molecules become anomalously broadened at the retardation level in a denaturing gradient, or double bands may appear in a uniform denaturant concentration. These properties are associated only with molecules for which the distribution of stability calculated by the Poland-Fixman-Freire algorithms indicates that the region of lowest stability does not extend to an end of the molecule. Retention of helicity at the ends is shown by the difference in the effect of base substitution in the end domains and in the least stable domain. Both the appearance of double bands and band broadening can be explained by invoking a hypothetical metastable intermediate in melting, which is converted into the equilibrium melted form at a relatively slow rate, depending on both denaturant concentration and field strength. A kinetic model permits plausible rate constants to be inferred from the patterns. Despite the increased band width, sequence variants with base changes in the least stable domain result in readily detectable band shifts in the gradient.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Abrams ES,Murdaugh SE,Lerman LS

doi

10.1093/nar/23.14.2775

subject

Has Abstract

pub_date

1995-07-25 00:00:00

pages

2775-83

issue

14

eissn

0305-1048

issn

1362-4962

pii

5j0127

journal_volume

23

pub_type

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