PMID:10572190
| Citation |
Singer, MJ, Podyminogin, MA, Metcalf, MA, Reed, MW, Brown, DA, Gamper, HB, Meyer, RB and Wydro, RM (1999) Targeted mutagenesis of DNA with alkylating RecA assisted oligonucleotides. Nucleic Acids Res. 27:e38 |
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| Abstract |
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. |
| Links | |
| Keywords |
Alkylating Agents/metabolism; Alkylation; Animals; Cell Line; Cloning, Molecular; DNA/genetics; DNA/metabolism; Escherichia coli/genetics; Escherichia coli/metabolism; Genes, Suppressor; Genetic Vectors; Mechlorethamine/metabolism; Mutagenesis, Site-Directed; Oligodeoxyribonucleotides/metabolism; Plasmids; RNA, Transfer/genetics; Rec A Recombinases/metabolism; Sequence Analysis, DNA |
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