Incorporation of 4′-C-aminomethyl-2′-O-methylthymidine into DNA by thermophilic DNA polymerases

Incorporation of 4'-C-aminomethyl-2'-O-methylthymidine into DNA by thermophilic DNA polymerases.

The dual modified nucleotide 4'-C-aminomethyl-2'-O-methylthymidine 5'-triphosphate was synthesized and enzymatically incorporated into DNA by the thermophilic DNA polymerases Pfu and Therminator III. The dual ribose modification imparted increased exonuclease resistance to DNA compared to the well-known 2'-O-methyl modification.

Incorporation of 40-C-aminomethyl-20-O-methylthymidine into DNA by thermophilic DNA polymerasesw

Functional nucleic acids such as deoxyribozymes (DNA enzymes) and aptamers are attracting interest as potential therapeutic tools. In order to impart drug-like properties to these molecules, judicious incorporation of chemical modifications is warranted. Site-specifically installed nucleoside modifications are often deleterious at key residues in the catalytic core of DNA enzymes or in binding ...

Abundant ribonucleotide incorporation into DNA by yeast replicative polymerases.

Measurements of nucleoside triphosphate levels in Saccharomyces cerevisiae reveal that the four rNTPs are in 36- to 190-fold molar excess over their corresponding dNTPs. During DNA synthesis in vitro using the physiological nucleoside triphosphate concentrations, yeast DNA polymerase epsilon, which is implicated in leading strand replication, incorporates one rNMP for every 1,250 dNMPs. Pol del...

Ara-C incorporation into DNA.

Effects of a modified dye-labeled nucleotide spacer arm on incorporation by thermophilic DNA polymerases.

The ability of eight commercially available thermophilic DNA polymerases to sequentially incorporate fluorescently labeled nucleotides sequentially was analyzed by a gel based primer extension assay. Cy5-dUTP or a variant nucleotide in which the linker had been lengthened by 14 atoms between the dye and the nucleobase were compared. We found that the Cy5-dUTP with a longer linker resulted in lo...