title: Targeted mutagenesis of DNA with alkylating RecA assisted oligonucleotides.
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.
authors.affiliation: Epoch Pharmaceuticals, Inc., 12277 134th Ct. NE, Redmond, WA 98052, USA. msinger@epochpharm.com
authors.forename: M J
authors.initials: MJ
authors.lastname: Singer
authors.forename: M A
authors.initials: MA
authors.lastname: Podyminogin
authors.forename: M A
authors.initials: MA
authors.lastname: Metcalf
authors.forename: M W
authors.initials: MW
authors.lastname: Reed
authors.forename: D A
authors.initials: DA
authors.lastname: Brown
authors.forename: H B
authors.initials: HB
authors.lastname: Gamper
authors.forename: R B
authors.initials: RB
authors.lastname: Meyer
authors.forename: R M
authors.initials: RM
authors.lastname: Wydro
mesh_terms: D000477:Alkylating Agents / Q000378:metabolism*; D000478:Alkylation; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016153:Genes, Suppressor; D005822:Genetic Vectors; D008466:Mechlorethamine / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D010957:Plasmids; D012343:RNA, Transfer / Q000235:genetics; D011938:Rec A Recombinases / Q000378:metabolism*; D017422:Sequence Analysis, DNA
chemical_list: D000477:Alkylating Agents; D009838:Oligodeoxyribonucleotides; C096275:supF tRNA; D008466:Mechlorethamine; D004247:DNA; D012343:RNA, Transfer; D011938:Rec A Recombinases