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Nucleic Acids Research, Vol 27, Issue 24 e38-e38, Copyright © 1999 by Oxford University Press

ARTICLES

Targeted mutagenesis of DNA with alkylating RecA assisted oligonucleotides

MJ Singer, MA Podyminogin, MA Metcalf, MW Reed, DA Brown, HB Gamper, RB Meyer and RM Wydro
Epoch Pharmaceuticals, Inc., 12277 134th Ct. NE, Redmond, WA 98052, USA

Site-specific mutation was demonstrated in a shuttle vector system using nitrogen mustard-conjugated oligodeoxyribonucleotides (ODNs). Plasmid DNA was modified in vitro by ODNs containing all four DNA bases in the presence of Escherichia coli RecA protein. Up to 50% of plasmid molecules were alkylated in the targeted region of the supF gene and mutations resulted upon replication in mammalian cells. ODNs conjugated with either two chlorambucil moieties or a novel tetrafunctional mustard caused interstrand crosslinks in the target DNA and were more mutagenic than ODNs that caused only monoadducts.
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