Force Spectroscopy of DNA and RNA

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1、Chapter2ForceSpectroscopyofDNAandRNA:StructureandKineticsfromSingle-MoleculeExperimentsRebeccaBoltEttlinger,MichaelAskvadSørensen,andLeneBroengOddershedeAbstractForcespectroscopyofindividualDNAandRNAmoleculesprovidesuniqueinsightsintothestructureandmechanicsoftheseforlifesoessentialmole

2、cules.ObservationsofDNAandRNAmoleculesoneatatimeprovidespatial,structural,andtemporalinformationthatiscomplementarytotheinformationobtainedbyclassicalensemblemethods.Single-moleculeforcespectroscopyhasbeenrealizedonlywithinthelastdecades,anditssuccessiscruciallyconnectedtothetechnologic

3、aldevelopmentthathasallowedsingle-moleculeresolution.ThischapterprovidesanintroductiontoinvitroforcespectroscopyofindividualDNAandRNAmoleculesincludingthemostcommonlyusedtechniques,thetheoryandmethodologynecessaryforunderstandingthedata,andtheexcitingresultsachieved.Threecommonlyusedsin

4、gle-moleculemethodsareemphasized:opticaltweezers,magnetictweezers,andnanoporeforcespectroscopy.ThetheoryofDNAstretchandtwistundertensionisdescribedalongwithrelatedexperimentalexamples.Newprinciplesforextractingkineticandthermodynamicinformationfromnonequilibriumdataareoutlined,andfurthe

5、rexamplesaregivenincludingtheopeningofDNAandRNAstructurestorevealtheirenergylandscape.Finally,futureperspectivesforforcespectroscopyofDNAandRNAareoffered.Contents2.1Introduction.................................................................................252.2CommonlyUsedMethods.....

6、.............................................................262.2.1OpticalTweezers...................................................................262.2.2MagneticTweezers.................................................................292.2.3NanoporeForceSpectroscopy.................

7、....................................29R.B.EttlingerL.B.Oddershede(*)TheNielsBohrInstitute,UniversityofCopenhagen,Copenhagen,Denmarke-mail:rebecca@nbi.dk;oddershede@nbi.dkM.A.SørensenTheDepartmentofBiology,UniversityofCopenhagen,Copenhagen,Denmarke-mail:mas@bio.ku.dkJ.KJEMSetal.