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RESEARCH ARTICLE The three-dimensional structure of in vitro

The three-dimensional structure of in vitro
reconstituted Xenopus laevis chromosomes
by EM tomography
Peter König & Michael B. Braunfeld & John W. Sedat &
David A. Agard
Received: 10 December 2006 /Revised: 16 January 2007 /Accepted: 17 January 2007
# Springer-Verlag 2007
Abstract We have studied the in vitro reconstitution of
sperm nuclei and small DNA templates to mitotic chroma-
tin in Xenopus laevis egg extracts by three-dimensional
(3D) electron microscopy (EM) tomography. Using specif-
ically developed software, the reconstituted chromatin was
interpreted in terms of nucleosomal patterns and the overall
chromatin connectivity. The condensed chromatin formed
from small DNA templates was characterized by aligned
arrays of packed nucleosomal clusters having a typical
10-nm spacing between nucleosomes within the same cluster
and a 30-nm spacing between nucleosomes in different
clusters. A similar short-range nucleosomal clustering was


Source: Agard, David - Department of Biochemistry and Biophysics, University of California at San Francisco


Collections: Biotechnology; Biology and Medicine