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Science 12 August 1994:
Vol. 265. no. 5174, pp. 959 - 962
DOI: 10.1126/science.8052855
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Articles

Science, Vol 265, Issue 5174, 959-962
Copyright © 1994 by American Association for the Advancement of Science

articles

High-specificity DNA cleavage agent: design and application to kilobase and megabase DNA substrates

PS Pendergrast, YW Ebright, and RH Ebright

Department of Chemistry, Rutgers University, New Brunswick, NJ 08855.

Strategies to cleave double-stranded DNA at specific DNA sites longer than those of restriction endonucleases (longer than 8 base pairs) have applications in chromosome mapping, chromosome cloning, and chromosome sequencing--provided that the strategies yield high DNA-cleavage efficiency and high DNA-cleavage specificity. In this report, the DNA-cleaving moiety copper:o-phenanthroline was attached to the sequence-specific DNA binding protein catabolite activator protein (CAP) at an amino acid that, because of a difference in DNA bending, is close to DNA in the specific CAP-DNA complex but is not close to DNA in the nonspecific CAP-DNA complex. The resulting CAP derivative, OP26CAP, cleaved kilobase and megabase DNA substrates at a 22-base pair consensus DNA site with high efficiency and exhibited no detectable nonspecific DNA-cleavage activity.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Mechanistic studies on DNA damage by minor groove binding copper-phenanthroline conjugates.
B. C. Bales, T. Kodama, Y. N. Weledji, M. Pitie, B. Meunier, and M. M. Greenberg (2005)
Nucleic Acids Res. 33, 5371-5379
   Abstract »    Full Text »    PDF »
Role of Oxidative Stress in the Action of Vanadium Phosphotyrosine Phosphatase Inhibitors. REDOX INDEPENDENT ACTIVATION OF NF-kappa B.
C. M. Krejsa, S. G. Nadler, J. M. Esselstyn, T. J. Kavanagh, J. A. Ledbetter, and G. L. Schieven (1997)
J. Biol. Chem. 272, 11541-11549
   Abstract »    Full Text »    PDF »


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