Brennan C A, Gumport R I
Nucleic Acids Res. 1985 Dec 20;13(24):8665-84. doi: 10.1093/nar/13.24.8665.
Self-complementary oligodeoxyribonucleotides containing the base analogues 2-aminopurine, 2,6-diaminopurine, N6-methyladenine, uracil, and 5-bromouracil were synthesized by a general method that allows incorporation of the analogues at specific positions. The method uses chemically synthesized partial sequences but circumvents the need for protected base analogues by incorporating their unprotected 3',5'-bisphosphate derivatives enzymatically. T4 RNA ligase was used to add the analogues to the oligodeoxyribonucleotides with yields from 54 to greater than 95 percent. Oligodeoxyribonucleotides were joined to the oligodeoxyribonucleotides containing the analogues at their 3'-termini in yields from 22 to 81 percent. The high yields obtained in these joinings suggest that RNA ligase should be of general use for the specific incorporation of other deoxyribonucleotide analogues into oligodeoxyribonucleotides. The oligodeoxyribonucleotides containing the base analogues were characterized by their mobilities during HPLC, nucleoside compositions, sequences, and thermal stabilities.
通过一种通用方法合成了含有碱基类似物2-氨基嘌呤、2,6-二氨基嘌呤、N6-甲基腺嘌呤、尿嘧啶和5-溴尿嘧啶的自互补寡脱氧核糖核苷酸，该方法允许在特定位置掺入这些类似物。该方法使用化学合成的部分序列，但通过酶促掺入其未保护的3',5'-双磷酸衍生物来避免对受保护碱基类似物的需求。使用T4 RNA连接酶将类似物添加到寡脱氧核糖核苷酸中，产率为54%至大于95%。寡脱氧核糖核苷酸在其3'-末端与含有类似物的寡脱氧核糖核苷酸连接，产率为22%至81%。这些连接中获得的高产率表明，RNA连接酶在将其他脱氧核糖核苷酸类似物特异性掺入寡脱氧核糖核苷酸中应具有普遍用途。含有碱基类似物的寡脱氧核糖核苷酸通过其在高效液相色谱（HPLC）中的迁移率、核苷组成、序列和热稳定性进行表征。
