Mechanical opening of DNA by micro manipulation and force measurements

Ulrich Bockelmann; B. Essevaz-Roulet; Philippe Thomen; François Heslot

doi:10.1016/S1631-0705(02)01342-7

Biophysique à l'échelle de la molécule unique/Single molecule biophysics

Mechanical opening of DNA by micro manipulation and force measurements
[Ouverture mécanique de la molécule d'ADN par micro-manipulation et mesure de force]

Ulrich Bockelmann ¹ ; B. Essevaz-Roulet ¹ ; Philippe Thomen ¹ ; François Heslot ¹

¹ Laboratoire de physique de la matière condensée, École normale supérieure, 24, rue Lhomond, 75005 Paris, France

Comptes Rendus. Physique, Volume 3 (2002) no. 5, pp. 585-594.

Résumé
Abstract

Dans cet article nous résumons une partie de notre travail sur l'ouverture mécanique de l'ADN. Nous avons préparé des constructions moléculaires qui nous permettent d'attacher les deux brins complémentaires d'une extrémité d'une molécule unique d'ADN du bactériophage λ, séparément à une lamelle de microscope et à une bille microscopique. Dans une première série d'expériences, un levier souple de verre, agissant comme capteur de force, est attaché à la bille et sa deflexion est mesurée avec un microscope optique. Dans une seconde série, nous utilisons un piège optique interférométrique pour capturer la bille et mesurer son déplacement avec une résolution nanométrique. L'échantillon est déplacé lentement par rapport au système de mesure de force, induisant une ouverture progressive de la double hélice. La force, mesurée pendant cette ouverture mécanique, montre une variation caractéristique qui est reliée à la séquence des paires de base de la molécule d'ADN.

In this paper we summarize part of our work on the mechanical unzipping of DNA. We have prepared molecular constructions which allow us to attach the two complementary strands of one end of a single DNA molecule of the bacteriophage λ separately to a glass microscope slide and a microscopic bead. In a first series of experiments, a soft microneedle acting as a force sensor is attached to the bead and its deflection is measured with an optical microscope. In a second series, we use an optical trapping interferometer to capture the bead and to measure its displacement to nm resolution. The sample is slowly displaced with respect to the force measurement device, leading to a progressive opening of the double helix. The force measured during this mechanical opening shows a characteristic variation which is related to the base pair sequence of the DNA molecule.

Reçu le : 2002-01-31
Accepté le : 2002-04-23
Publié le : 2002-01-01

DOI : 10.1016/S1631-0705(02)01342-7

Keywords: micromanipulation, optical tweezers, DNA unzipping
Mot clés : micromanipulation, pinces optiques, ouverture de l'ADN

Affiliations des auteurs :

Ulrich Bockelmann ¹ ; B. Essevaz-Roulet ¹ ; Philippe Thomen ¹ ; François Heslot ¹

¹ Laboratoire de physique de la matière condensée, École normale supérieure, 24, rue Lhomond, 75005 Paris, France

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     title = {Mechanical opening of {DNA} by micro manipulation and force measurements},
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Ulrich Bockelmann; B. Essevaz-Roulet; Philippe Thomen; François Heslot. Mechanical opening of DNA by micro manipulation and force measurements. Comptes Rendus. Physique, Volume 3 (2002) no. 5, pp. 585-594. doi : 10.1016/S1631-0705(02)01342-7. https://comptes-rendus.academie-sciences.fr/physique/articles/10.1016/S1631-0705(02)01342-7/

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