Masud M M, Ozaki-Nakamura A, Satou F, Ohbayashi T, Ozaki H, Sawai H
Department of Applied Chemistry, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan.
Nucleic Acids Res Suppl. 2001(1):21-2. doi: 10.1093/nass/1.1.21.
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.
我们合成了多种在C5-α位带有亚甲基连接基的C5-取代2'-脱氧尿苷衍生物的5'-三磷酸酯。我们研究了C5-取代的2'-脱氧尿苷5'-三磷酸酯（dUTP）是否能作为通过聚合酶链反应（PCR）进行修饰DNA合成的底物。我们发现，只有来自极端嗜热古菌的耐热DNA聚合酶KOD dash，能接受修饰后的底物替代三磷酸胸苷（TTP）用于PCR，从而形成相应的修饰DNA。另一方面，没有其他DNA聚合酶能接受这些TTP类似物。
