Sequence-specific alkylation of dsDNA with derivatives of pyrimidine oligonucleotides conjugated to 2-chloroethylamine groups.
Reaction of homopyrimidine oligonucleotides bearing a 5'-terminal alkylating aromatic 2-chloroethyl-amino group with a bovine papilloma vector expressing human interferon-gamma was investigated. The oligonucleotide derivatives bound to corresponding homopurine-homopyrimidine sequences in dsDNA and alkylated guanosine residues at these sites in the purine strand of the target. The alkylated DNA can be cleaved at the modified residues. At pH 5.4, the reaction was highly specific to the target sequences; at pH less than 5, some nonspecific reactions were observed at the sequences partially complementary to the oligonucleotides. Elongation of the linker between the alkylating group and the oligonucleotide phosphate increased the alkylation efficiency. Repeated treatment of the DNA with gradually increased concentrations of the reagent resulted in quantitative modification of the target guanosines.
Brossalina
E B
EB
Institute of Clinical Immunology, Siberian Division, USSR Academy of Medical Sciences, Novosibirsk.
Demchenko
E N
EN
Vlassov
V V
VV
Mamaev
S V
SV
D000478:Alkylation; D001483:Base Sequence; D052688:Bovine papillomavirus 1 / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005021:Ethylamines; D005822:Genetic Vectors; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007371:Interferon-gamma / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000378:metabolism*; D015183:Restriction Mapping
D005021:Ethylamines; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D007371:Interferon-gamma; D004247:DNA; C010076:2-chloroethylamine