Kuwahara Masayasu, Hososhima Shin-ichi, Takahata Yumi, Kitagata Rina, Shoji Atsushi, Hanawa Kazuo, Ozaki Akiko N, Ozaki Hiroaki, Sawai Hiroaki
Department of Applied Chemistry, Gunma University, Kiryu, Gunma 376-8515, Japan.
Nucleic Acids Res Suppl. 2003(3):37-8. doi: 10.1093/nass/3.1.37.
Modified analogs of 2'-deoxycytidine triphosphates bearing (6-aminohexyl)carbamoylmethyl or 7-amino-2,5-dioxaheptyl linker at a C5 position were designed and synthesized. Both analogs were found to be good substrates for Vent(exo-) DNA polymerase during PCR, resulting in the corresponding full-length modified DNAs, respectively. Moreover, we have demonstrated simultaneous incorporation of three different modified nucleotides into a DNA strand by PCR using triphosphates of 5-(3-aminopropynyl)dUTP, 5-[(6-aminohexyl)carbamoylmethyl]dCTP and 2-amino-dATP (dDTP) or N6-methyl-dATP in place of the natural nucleoside triphosphates TTP, dCTP and dATP.
设计并合成了在C5位置带有（6-氨基己基）氨基甲酰甲基或7-氨基-2,5-二氧杂庚基连接基的2'-脱氧胞苷三磷酸的修饰类似物。发现这两种类似物在PCR过程中都是Vent（exo-）DNA聚合酶的良好底物，分别产生相应的全长修饰DNA。此外，我们已经证明，通过使用5-（3-氨基丙炔基）dUTP、5-[(6-氨基己基)氨基甲酰甲基]dCTP和2-氨基-dATP（dDTP）或N6-甲基-dATP的三磷酸代替天然核苷三磷酸TTP、dCTP和dATP，通过PCR可将三种不同的修饰核苷酸同时掺入DNA链中。
