28 research outputs found
Gel Electrophoresis of DNA Knots in Weak and Strong Electric Fields
Gel electrophoresis allows to separate knotted DNA (nicked circular) of equal
length according to the knot type. At low electric fields, complex knots being
more compact, drift faster than simpler knots. Recent experiments have shown
that the drift velocity dependence on the knot type is inverted when changing
from low to high electric fields. We present a computer simulation on a lattice
of a closed, knotted, charged DNA chain drifting in an external electric field
in a topologically restricted medium. Using a simple Monte Carlo algorithm, the
dependence of the electrophoretic migration of the DNA molecules on the type of
knot and on the electric field intensity was investigated. The results are in
qualitative agreement with electrophoretic experiments done under conditions of
low and high electric fields: especially the inversion of the behavior from low
to high electric field could be reproduced. The knot topology imposes on the
problem the constrain of self-avoidance, which is the final cause of the
observed behavior in strong electric field.Comment: 17 pages, 5 figure
Polarization-dependence of anomalous scattering in brominated DNA and RNA molecules, and importance of crystal orientation in single- and multiple-wavelength anomalous diffraction phasing
In this paper the anisotropy of anomalous scattering at the Br K-absorption edge in brominated nucleotides is investigated, and it is shown that this effect can give rise to a marked directional dependence of the anomalous signal strength in X-ray diffraction data. This implies that choosing the correct orientation for crystals of such molecules can be a crucial determinant of success or failure when using single- and multiple-wavelength anomalous diffraction (SAD or MAD) methods to solve their structure. In particular, polarized absorption spectra on an oriented crystal of a brominated DNA molecule were measured, and were used to determine the orientation that yields a maximum anomalous signal in the diffraction data. Out of several SAD data sets, only those collected at or near that optimal orientation allowed interpretable electron density maps to be obtained. The findings of this study have implications for instrumental choices in experimental stations at synchrotron beamlines, as well as for the development of data collection strategy programs
Exploiting X-ray induced anisotropic lattice changes to improve intensity extraction in protein powder diffraction: Application to heavy atom detection
X-ray induced anisotropic variations of cell parameters in porcine pancreatic elastase (PPE) were used in a multi-Pawley refinement in order to improve the deconvolution of overlapping peaks occurring in the high-angle region of the powder pattern. The benefit of combining scans is demonstrated by an improvement in the quality of the isomorphous difference Patterson maps used to detect the positions of heavy atoms in a uranyl derivative of PPE