High-Specificity DNA Cleavage Agent: Design and Application to Kilobase and Megabase DNA Substrates
Copyright policy )High-Specificity DNA Cleavage Agent: Design and Application to Kilobase and Megabase DNA Substrates
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, OP 26 CAP, 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.
- Rutgers, The State University of New Jersey United States
DNA-Binding Proteins, Cyclic AMP Receptor Protein, Base Sequence, Molecular Structure, Molecular Sequence Data, Nucleic Acid Conformation, DNA, Phenanthrolines
DNA-Binding Proteins, Cyclic AMP Receptor Protein, Base Sequence, Molecular Structure, Molecular Sequence Data, Nucleic Acid Conformation, DNA, Phenanthrolines
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