Brázdilová Petra, Vrábel Milan, Pohl Radek, Pivonková Hana, Havran Ludek, Hocek Michal, Fojta Miroslav
Institute of Biophysics, v.v.i. Academy of Sciences of the Czech Republic, Kralovopolska 135, 61265 Brno, Czech Republic.
Chemistry. 2007;13(34):9527-33. doi: 10.1002/chem.200701249.
Modified dATP (2'-deoxyadenosine-5'-triphosphate) and dUTP (2'-deoxyuridine-5'-triphosphate) bearing ferrocene (Fc) labels linked via a conjugate acetylene spacer were prepared by single-step aqueous-phase cross-coupling reactions of 7-iodo-7-deaza-dATP or 5-iodo-dUTP with ethynylferrocene. The Fc-labeled dNTPs were good substrates for DNA polymerases and were efficiently incorporated to DNA by primer extension (PEX). Electrochemical analysis of the 2'-deoxyribonucleoside triphosphates (dNTPs) and PEX products revealed significant differences in redox potentials of the Fc label bound either to U or to 7-deazaA and between isolated dNTPs and conjugates incorporated to DNA. Prospective bioanalytical applications are outlined.
通过7-碘-7-脱氮-dATP或5-碘-dUTP与乙炔基二茂铁的单步水相交叉偶联反应，制备了带有通过共轭乙炔间隔基连接的二茂铁（Fc）标签的修饰dATP（2'-脱氧腺苷-5'-三磷酸）和dUTP（2'-脱氧尿苷-5'-三磷酸）。Fc标记的dNTP是DNA聚合酶的良好底物，并通过引物延伸（PEX）有效地掺入到DNA中。对2'-脱氧核糖核苷三磷酸（dNTP）和PEX产物的电化学分析表明，与U或7-脱氮A结合的Fc标签的氧化还原电位以及分离的dNTP与掺入DNA的缀合物之间存在显著差异。概述了潜在的生物分析应用。
