Enzymatic synthesis of modified DNA by PCR.
We synthesized various 5'-triphosphates of C5-substituted 2'-deoxyuridine derivatives bearing methylene linker at C5-alpha position. We examined whether the C5-substituted 2'-deoxyuridine 5'-triphosphates (dUTP) can work as a substrate for the modified DNA synthesis by PCR. We found that only KOD dash DNA polymerase, a thermostable DNA polymerase from extremely thermophilic archaeum, accepted the modified substrates in place of TTP for PCR forming the corresponding modified DNAs. On the other hand, no other DNA polymerase could accept these TTP analogues.
Masud
M M
MM
Department of Applied Chemistry, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan.
Ozaki-Nakamura
A
A
Satou
F
F
Ohbayashi
T
T
Ozaki
H
H
Sawai
H
H
D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000138:chemical synthesis / Q000737:chemistry; D009838:Oligodeoxyribonucleotides / Q000096:biosynthesis* / Q000737:chemistry; D016133:Polymerase Chain Reaction
D003856:Deoxyuracil Nucleotides; D009838:Oligodeoxyribonucleotides; C027078:deoxyuridine triphosphate; D004247:DNA; D004259:DNA-Directed DNA Polymerase