Gene Manipulation using Laser

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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.