Tauraitė Daiva, Jakubovska Jevgenija, Dabužinskaitė Julija, Bratchikov Maksim, Meškys Rolandas
Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Sauletekio al. 7, Vilnius LT-10257, Lithuania.
Department of Physiology, Biochemistry, Microbiology and Laboratory Medicine, Faculty of Medicine, Vilnius University, M. K. Čiurlionio g. 21, Vilnius LT-03101, Lithuania.
Molecules. 2017 Apr 22;22(4):672. doi: 10.3390/molecules22040672.
The synthesis of novel modified nucleotides and their incorporation into DNA sequences opens many possibilities to change the chemical properties of oligonucleotides (ONs), and, therefore, broaden the field of practical applications of modified DNA. The chemical synthesis of nucleotide derivatives, including ones bearing thio-, hydrazino-, cyano- and carboxy groups as well as 2-pyridone nucleobase-containing nucleotides was carried out. The prepared compounds were tested as substrates of terminal deoxynucleotidyl transferase (TdT). The nucleotides containing ⁴-aminocytosine, 4-thiouracil as well as 2-pyridone, 4-chloro- and 4-bromo-2-pyridone as a nucleobase were accepted by TdT, thus allowing enzymatic synthesis of 3'-terminally modified ONs. The successful UV-induced cross-linking of 4-thiouracil-containing ONs to TdT was carried out. Enzymatic post-synthetic 3'-modification of ONs with various photo- and chemically-reactive groups opens novel possibilities for future applications, especially in analysis of the mechanisms of polymerases and the development of photo-labels, sensors, and self-assembling structures.
新型修饰核苷酸的合成及其掺入DNA序列为改变寡核苷酸（ONs）的化学性质开辟了多种可能性，从而拓宽了修饰DNA的实际应用领域。进行了核苷酸衍生物的化学合成，包括带有硫基、肼基、氰基和羧基的衍生物以及含2-吡啶酮核碱基的核苷酸。所制备的化合物作为末端脱氧核苷酸转移酶（TdT）的底物进行了测试。含有⁴-氨基胞嘧啶、4-硫尿嘧啶以及作为核碱基的2-吡啶酮、4-氯-和4-溴-2-吡啶酮的核苷酸被TdT接受，从而允许酶促合成3'-末端修饰的ONs。成功地进行了含4-硫尿嘧啶的ONs与TdT的紫外诱导交联。用各种光反应性和化学反应性基团对ONs进行酶促合成后3'-修饰为未来应用开辟了新的可能性，特别是在聚合酶机制分析以及光标签、传感器和自组装结构的开发方面。
