Nucleic Acids Research, Vol 24, Issue 13 2483-2487, Copyright © 1996 by Oxford University Press
AG Veselkov, VV Demidov, PE Nielson and MD Frank-Kamenetskii
Although significant efforts have been directed at developing efficient
techniques for rare and super rare genome cutting, only limited success has
been achieved. Here we propose a new approach to solve this problem. We
demonstrate that peptide nucleic acid 'clamps' (bis-PNAs) bind strongly and
sequence specifically to short homopyrimidine sites on lambda and yeast
genomic DNAs. Such binding efficiently shields methylation/restriction
sites which overlap with the bis-PNA binding sites from enzymatic
methylation. After removing the bis-PNA, the genomic DNAs are
quantitatively cleaved by restriction enzymes into a limited number of
pieces of lengths from several hundred kbp to several Mbp. By combining
various bis-PNAs with different methylation/restriction enzyme pairs, a
huge new class of genome rare cutters can be created. These cutters cover
the range of recognition specificities where very few, if any, cutters are
now available.
ARTICLES
A new class of genome rare cutters
Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, 36 Cummington Street, Boston, MA 02215, USA.
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