Lebedeva N A, Rechkunova N I, Dezhurov S V, Degtiarev S Kh, Lavrik O I
Bioorg Khim. 2004 Jul-Aug;30(4):369-74. doi: 10.1023/b:rubi.0000037258.51646.76.
Substrate properties of several dTTP analogues bearing a photoreactive 2-nitro-5-azidobenzoyl (NAB) group attached at position 5 of uracil through linkers of various lengths, dTTP-NAB-x-dUTP (where x = 2, 4, 7-13 is the number of atoms in the linker), were studied. All the analogues are substrates for thermostable Thermus thermophilus B35 DNA polymerase in the elongation reaction of the 5'-32P-labeled primer-template complex. The kinetic parameters of some of the analogues were determined and compared with those of natural dTTP. It was shown that an increase in the linker length results in a higher efficiency of the analogue. The incorporation of NAB-x-dUMP residues into the 3'-primer end did not impede a further elongation of the chain in the presence of natural dNTP.
研究了几种在尿嘧啶5位通过不同长度连接子连接有光反应性2-硝基-5-叠氮苯甲酰基（NAB）基团的dTTP类似物，即dTTP-NAB-x-dUTP（其中x = 2、4、7 - 13为连接子中的原子数）的底物性质。在5'-32P标记的引物-模板复合物的延伸反应中，所有类似物都是嗜热栖热菌B35耐热DNA聚合酶的底物。测定了部分类似物的动力学参数，并与天然dTTP的动力学参数进行比较。结果表明，连接子长度增加会导致类似物效率提高。在天然dNTP存在的情况下，将NAB-x-dUMP残基掺入3'-引物末端并不妨碍链的进一步延伸。
