Nallagatla Subba Rao, Heuberger Benjamin, Haque Azizul, Switzer Christopher
Department of Chemistry, University of California, Riverside, California 92521, USA.
J Comb Chem. 2009 May-Jun;11(3):364-9. doi: 10.1021/cc800178m.
A library of all possible substitutions of guanine by iso-guanine (iG) in the thrombin aptamer was prepared by split and mix synthesis. A colorimetric assay was used to screen for functional oligomers in the library. Colorimetrically active oligonucleotides were selected and sequenced by the Maxam-Gilbert method. The sequenced oligonucleotides were individually resynthesized, and their affinities for thrombin were assayed by isothermal titration calorimetry. Three aptamer sequences containing iG were found to have enhanced binding activity to human alpha-thrombin compared to the parent aptamer.
通过拆分混合合成法构建了凝血酶适体中鸟嘌呤被异鸟嘌呤（iG）全部可能替代的文库。采用比色法筛选文库中的功能性寡聚体。选择具有比色活性的寡核苷酸并通过Maxam-Gilbert法进行测序。对测序后的寡核苷酸进行单独重新合成，并通过等温滴定量热法测定它们对凝血酶的亲和力。发现与亲本适体相比，三个含有iG的适体序列对人α-凝血酶具有增强的结合活性。
