Gao H, Yang M, Patel R, Cook A F
PharmaGenics, Inc., Allendale, NJ 07401, USA.
Nucleic Acids Res. 1995 Jun 11;23(11):2025-9. doi: 10.1093/nar/23.11.2025.
An effective, convenient method for the circularization of oligonucleotides has been developed. This procedure involved preparation of an oligonucleotide with backbone-linked 5'- and 3'-terminal hexamethylenethiol groups, followed by oxidation of the thiol groups with air of oxygen to produce the corresponding circular sequence bridged via a bis(hexamethylene)-disulfide moiety. The method has been applied to the circularization of oligodeoxynucleotide sequences of varying lengths (5, 10, 15, 20, 30 and 40 bases), and the circularization process was highly efficient as shown by HPLC or gel electrophoresis of the crude reaction mixtures. Competing reactions such as dimerization were not significant except for the longer sequences (30 and 40 bases). The circularization of an eight base RNA sequence was also accomplished, as well as hexa-ethylene glycol bridged poly-T sequences capable of triplex formation.
已开发出一种有效、便捷的寡核苷酸环化方法。该过程包括制备带有与主链相连的5'-和3'-末端六亚甲基硫醇基团的寡核苷酸，然后用空气或氧气氧化硫醇基团，以产生通过双（六亚甲基）-二硫部分桥接的相应环状序列。该方法已应用于不同长度（5、10、15、20、30和40个碱基）的寡脱氧核苷酸序列的环化，粗反应混合物的高效液相色谱（HPLC）或凝胶电泳表明环化过程效率很高。除了较长的序列（30和40个碱基）外，诸如二聚化等竞争反应并不显著。还完成了一个八个碱基的RNA序列的环化，以及能够形成三链体的六乙二醇桥连的聚T序列的环化。
