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2‐Allyl‐ and Propargylamino‐dATPs for Site‐Specific Enzymatic Introduction of a Single Modification in the Minor Groove of DNA
Author(s) -
Matyašovský Ján,
Pohl Radek,
Hocek Michal
Publication year - 2018
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201803973
Subject(s) - minor groove , dna , oligonucleotide , nucleotide , primer extension , chemistry , primer (cosmetics) , biochemistry , stereochemistry , microbiology and biotechnology , combinatorial chemistry , biology , base sequence , gene , organic chemistry
A series of 2‐alkylamino‐2′‐deoxyadenosine triphosphates (dATP) was prepared and found to be substrates for the Therminator DNA polymerase, which incorporated only one modified nucleotide into the primer. Using a template encoding for two consecutive adenines, conditions were found for incorporation of either one or two modified nucleotides. In all cases, addition of a mixture of natural dNTPs led to primer extension resulting in site‐specific single modification of DNA in the minor groove. The allylamino‐substituted DNA was used for the thiol‐ene addition, whereas the propargylamino‐DNA for the CuAAC click reaction was used to label the DNA with a fluorescent dye in the minor groove. The approach was used to construct FRET probes for detection of oligonucleotides.