Chemical synthesis of novel base pairs and their enzymatic incorporation into DNA.
A novel base pair, 2-amino-6-(N,N-dimethylamino)purine (denoted x) and the counter part, pyridin-2-one (denoted y) were designed. The bulky 6-dimethylamino group of x is expected to eliminate base pairing with all natural bases. The phosphoramidite of x for DNA templates and the 2'-deoxyribonucleoside triphosphate of y (dyTP) for a substrate were synthesized, and the selectivity of the enzymatic incorporation of dyTP opposite x in the templates was examined. dyTP was preferentially incorporated opposite x than canonical dNTPs by Klenow fragment of Escherichia coli DNA polymerase I. While dyTP was also incorporated opposite A and G, the misincorporation was suppressed in the presence of dTTP and dCTP, respectively.
Ishikawa
M
M
Yokoyama CytoLogic Project, The RIKEN Institute, Saitama, Japan.
Hirao
I
I
Yokoyama
S
S
D020029:Base Pairing; D001483:Base Sequence; D004247:DNA / Q000138:chemical synthesis*; D004256:DNA Polymerase I; D017931:DNA Primers; D003854:Deoxyribonucleotides / Q000138:chemical synthesis; D015195:Drug Design; D004926:Escherichia coli / Q000201:enzymology; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis*; D011687:Purines / Q000138:chemical synthesis*; D011728:Pyridones / Q000138:chemical synthesis*; D013698:Templates, Genetic
D017931:DNA Primers; D003854:Deoxyribonucleotides; D009838:Oligodeoxyribonucleotides; D011687:Purines; D011728:Pyridones; D004247:DNA; D004256:DNA Polymerase I