[Construction of DNA-probes and their immunochemical detection using nonradioactive hybridization tests].
Simple and efficient chemical approaches to preparation of DNA probes carrying 2,4-dinitrophenyl, dansyl or biotin residues were developed. The residues were introduced using following DNA derivatization procedures: a) transamination of cytidine residues with O-(4-aminobutyl)hydroxylamine; b) mercuration of pyrimidine residues followed by beta-mercaptoethanol modification. It was shown that 2,4-dinitrophenyl-containing DNA probes can be used for nonradioactive hybridization detection of nucleic acids. DNP-DNA: DNA complexes were detected using mouse antibodies specific to 2,4-dinitrophenyl groups, which were developed with peroxidase-conjugated antimouse immunoglobulins. Peroxidase-catalyzed chemoluminescent reaction of luminol oxidation with hydrogen peroxide allowed to detect 10 picograms of the dinitrophenylated single-stranded DNA probe.
Brosalina
E B
EB
Vlasov
V V
VV
Grachev
S A
SA
Demchenko
E N
EN
D001710:Biotin; D055598:Chemical Phenomena; D002621:Chemistry; D004247:DNA / Q000276:immunology; D003619:Dansyl Compounds; D004136:Dinitrobenzenes; D006241:Haptens; D009693:Nucleic Acid Hybridization; D011089:Polydeoxyribonucleotides / Q000138:chemical synthesis
D003619:Dansyl Compounds; D004136:Dinitrobenzenes; D006241:Haptens; D011089:Polydeoxyribonucleotides; D001710:Biotin; D004247:DNA
Konstruirovanie DNK-zondov i ikh immunokhimicheskoe opredelenie v neradioaktivnykh gibridizatsionnykh testakh.