Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov Street, 119991 Moscow, Russia.
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov Street, 119991 Moscow, Russia.
Bioorg Chem. 2020 Jun;99:103829. doi: 10.1016/j.bioorg.2020.103829. Epub 2020 Apr 8.
Deoxyuridine triphosphate derivatives (dUTPs) modified at the C5 position of the pyrimidine ring with various aromatic hydrocarbon substituents of different hydrophilicities have been synthesized. The aromatic hydrocarbon substituents were attached to dUTPs via a CHCHCHNHCOCH linker. The efficiency of the PCR incorporation of modified dUMPs using Taq, Tth, Vent (exo-) and Deep Vent (exo-) polymerases and a model DNA template containing one, two and three adjacent adenine nucleotides at three different sites within the sequence was investigated. For all the polymerases used, the yield of the modified PCR product was significantly increased with increasing hydrophilicity of the aromatic hydrocarbon substituent. In particular, for the above polymerases, the efficiency of the incorporation of dUMPs modified with the most hydrophilic of the studied aromatic hydrocarbon substituents, a 4-hydroxyphenyl residue, was 60-85% of the efficiency of dTMP incorporation. At the same time, the relative efficiencies of the incorporation of dUMPs modified with 2-, 4-methoxyphenyl, phenyl and 4-nitrophenyl substituents ranged from 20 to 50% and were 2-18% for the 1-naphthalene and 4-biphenyl groups, which were the most hydrophobic of the studied aromatic hydrocarbon substituents.
已合成了嘧啶环 C5 位带有各种不同亲水性芳族烃取代基的脱氧尿苷三磷酸衍生物（dUTP）。芳族烃取代基通过 CHCHCHNHCOCH 接头连接到 dUTP 上。使用 Taq、Tth、Vent（外切）和 Deep Vent（外切）聚合酶以及含有一个、两个和三个相邻腺嘌呤核苷酸的模型 DNA 模板，研究了在序列中的三个不同位置，研究了修饰的 dUMP 对 PCR 掺入的效率。对于所有使用的聚合酶，随着芳族烃取代基亲水性的增加，修饰的 PCR 产物的产率显著增加。特别是对于上述聚合酶，与研究的芳族烃取代基中最亲水的 4-羟基苯基残基修饰的 dUMP 的掺入效率是 dTMP 掺入效率的 60-85%。同时，用 2-、4-甲氧基苯基、苯基和 4-硝基苯基取代基修饰的 dUMP 的相对掺入效率范围为 20-50%，而 1-萘基和 4-联苯基基团的相对掺入效率为 2-18%，它们是研究的芳族烃取代基中最疏水的基团。
