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  • 1
    Digitale Medien
    Digitale Medien
    College Park, Md. : American Institute of Physics (AIP)
    The Journal of Chemical Physics 114 (2001), S. 5049-5060 
    ISSN: 1089-7690
    Quelle: AIP Digital Archive
    Thema: Physik , Chemie und Pharmazie
    Notizen: We considered an irreversible biochemical intrachain reaction of supercoiled DNA as a random event that occurs, with some probability, at the instant of collision between two reactive groups attached to distant sites of the DNA molecule. For sufficiently small intrinsic rate constant kI, the dominant process contributing to the productive collisions is the quasione-dimensional reptation of the strands forming the superhelix. The mean reaction time is then given by τF+1/kIcL, where τF is the mean time of the first collision caused by reptation, and cL is the local concentration of one reactive group around the other. The internal reptation of DNA strands was simulated by the repton model, in which a superhelix branch is approximated by a string of beads placed in a row of cells. This simple model allows semiquantitative estimation of τF and cL (in some arbitrary units) for a large range of the DNA lengths L. The repton chain was calibrated with the help of the data available for small supercoiled plasmids from Monte Carlo and Brownian dynamics simulations. The repton model and the Brownian dynamics give the same form of the distribution of the first collision time. Our estimations show that, for opposite sites of the chain, the mean first collision time τF varies from 5 ms (L=2.5 kb) to 1 s (L=20 kb). The corresponding cL values (for the reaction radius 10 nm) are 3×10−6 and 2×10−7 M. © 2001 American Institute of Physics.
    Materialart: Digitale Medien
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  • 2
    ISSN: 1573-6857
    Schlagwort(e): DNA superhelix diameter ; Light scattering ; SANS
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We present here recent results on the structure of superhelical DNA and its changes with salt concentration between 0.01 and 1.5 M NaCl. Scattering curves of two different superhelical DNAs were determined by static light scattering. The measured radii of gyration do not change significantly with salt concentration. Small-angle neutron scattering, together with calculations from a Monte Carlo model, allows to determine the superhelix diameter. Measured and simulated scattering curves agreed almost quantitatively. Experimentally we find that the diameter decreases from 16.0±0.9 nm at 10 mM to 9.0±0.7 nm at 100 mM NaCl. The superhelix diameter from the simulated conformations decreased from 18.0±1.5 nm at 10 mM to 9.4±1.5 nm at 100 mM NaCl. At higher salt concentrations up to 1.5 M NaCl, the diameter stays constant at 9 nm.
    Materialart: Digitale Medien
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  • 3
    ISSN: 0006-3525
    Schlagwort(e): Chemistry ; Polymer and Materials Science
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Chemie und Pharmazie
    Notizen: Using dynamic light scattering (DLS), we have measured the diffusion coefficient and internal motions of narrow topoisomer distributions of pUCl8 DNA (2687 base pairs) as a function of linking number. The topoisomer distributions were prepared by C-18 reversed phase high performance liquid chromatography separation of topoisomerase I/ethidium-generated topoisomer families. The measured diffusion coefficients agree well with those predicted by a Monte Carlo model for the generation of equilibrium ensembles of DNA topoisomers of defined linking number. The only parameters used in the model were the bending persistence length, torsional rigidity, and hydrodynamic radius of DNA known independently from other techniques. Two different values for the torsional rigidity, α = 4 · 10-12 and 8.8 · 10-12 dyn cm, were used. Intrachain interactions in the DNA were taken into account by using an “effective DNA radius,” which was varied between rDNA = 1-8 nm. The best agreement between the measured and calculated values was obtained for α = 4 · 10-12 dyn cm, rDNA = 4 nm. The internal motions of the DNA topoisomers were characterized by the amplitude of the fast relaxation of the DLS autocorrelation function. Our earlier result that supercoiling leads to a decrease in the amplitude of internal motion was confirmed. In addition, we see a characteristic maximum of the internal motion amplitude at a superhelix density of σ = -0.03. The maximum occurs in the same range as a structural transition in pUC8 dimers previously described by L. Song et al. (1990 J. Mol. Biol. 214, 307-326). © 1994 John Wiley & Sons, Inc.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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