Gene Manipulation using Laser

我们以5-碘脱氧尿苷为原料，高产率地合成了各种乙烯基尿嘧啶类似物。乙烯基尿嘧啶被转化为亚磷酰胺形式，然后掺入到寡核苷酸(ODN)的5‘端。利用含有乙烯基尿嘧啶的ODN，在3‘端含有嘧啶碱基的ODN和模板ODN的存在下进行光配基。用聚丙烯酰胺凝胶电泳法测定其光催化效率。含羧基的嘧啶类化合物的光配位反应活性是5-乙烯基尿苷的10倍。我们还找到了一条合成乙烯基尿嘧啶类似物的有效路线。在制备了含有5-甲酰基尿嘧啶的ODN后，通过Wittig反应合成了具有乙烯基尿嘧啶类似物的ODN。

We synthesized various vinyluracil analogs from 5-iododeoxyuridine-in a high yield. Vinyluracils were converted to the phosphoramidite form, and then incorporated to 5' end of oligodeoxynucleotides (ODNs). Using the vinyluracil-containing ODN, photoligation was carried out in the presence of the ODN containing pyrimidine base at 3' end and the template ODN. The photoligation efficiency was quantified using polyacrylamide gel electrophoresis. The pyrimidine possessing a carboxyvinyl group exhibited 10 times higher photoligation reactivity than 5-vinyluridine. We also found an efficient synthetic route for vinyluracil analogs. After the ODN containing 5-formyluracil was prepared, ODNs possessing vinyluracil analogs were synthesized using Wittig reaction for 5-formyluracil.