Barriers against DNA-loop formation in a porous matrix

Björn Åkerman
Phys. Rev. E 54, 6685 – Published 1 December 1996
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Abstract

The resistance towards DNA bending imposed by a porous matrix has been investigated by studying the rate of helix-loop penetration through agarose gel pores as driven by electric fields between 1.1 and 6.7 V/cm. YOYO-stained DNA molecules (680 kilo-base-pairs) were prepared in a well-defined globally oriented state by an electrophoretic procedure (YOYO denotes dimer of oxazole-yellow). Loop initiation by a field perpendicular to the global orientation was detected by liner dichroism (LD) spectroscopy in terms of an initial net helix orientation perpendicular to the applied field direction, reflecting the stretching of the chain between the loopholes by the initial growth of the comparatively weakly oriented loop heads. The rate of loop nucleation exhibits a strong field dependence in agreement with a model based on the entropy cost of loop formation. The effect of increasing the average pore radius from 0.7 to 3 P, where P is the persistence length of DNA (500 Å), is significantly weaker than predicted from the model, however. After initially being perpendicular, the net helix orientation is eventually along the field direction, and during this phase the LD exhibits several oscillations before reaching a steady state. By comparison with fluorescence microscopy observations on individual molecules under identical conditions the LD oscillations are identified in terms of loop growth and competition. The spectroscopically measured average rates of these later loop processes exhibit considerably weaker field dependence than loop nucleation, and with power-law dependencies (E1.22) in agreement with the DNA coils being stretched by electrophoretic transport of the polymer ends.

  • Received 15 April 1996

DOI:https://doi.org/10.1103/PhysRevE.54.6685

©1996 American Physical Society

Authors & Affiliations

Björn Åkerman*

  • Department of Physical Chemistry, Chalmers University of Technology, Göteborg 41296, Sweden

  • *FAX: +46317723858. Electronic address: baa@phc.chalmers.se

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Vol. 54, Iss. 6 — December 1996

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